CXCR3 antagonists

ABSTRACT

Compounds, compositions and methods that are useful in the treatment of inflammatory and immune conditions and diseases are provided herein. In particular, the invention provides compounds which modulate the expression and/or function of a chemokine receptor. The subject methods are useful for the treatment of inflammatory and immunoregulatory disorders and diseases, such as multiple sclerosis, rheumatoid arthritis and type I diabetes.

CROSS-REFERENCES TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.60/296,499 filed Jun. 6, 2001, which is incorporated by reference hereinin its entirety.

BACKGROUND OF THE INVENTION

Chemokines are chemotactic cytokines that are released by a wide varietyof cells to attract macrophages, T cells, eosinophils, basophils andneutrophils to sites of inflammation (reviewed in Schall, Cytokine,3:165-183 (1991), Schall, et al., Curr. Opin. Immunol., 6:865-873 (1994)and Murphy, Rev. Immun., 12:593-633 (1994)). In addition to stimulatingchemotaxis, other changes can be selectively induced by chemokines inresponsive cells, including changes in cell shape, transient rises inthe concentration of intracellular free calcium ions ([Ca²⁺])₁, granuleexocytosis, integrin upregulation, formation of bioactive lipids (e.g.,leukotrienes) and respiratory burst, associated with leukocyteactivation. Thus, the chemokines are early triggers of the inflammatoryresponse, causing inflammatory mediator release, chemotaxis andextravasation to sites of infection or inflammation.

There are four classes of chemokines, CXC (α), CC (β), C(γ), and CX₃C(δ), depending on whether the first two cysteines are separated by asingle amino acid (C-X-C), are adjacent (C-C), have a missing cysteinepair (C), or are separated by three amino acids (CXC₃). Theα-chemokines, such as interleukin-8 (IL-8), melanoma growth stimulatoryactivity protein (MGSA), and stromal cell derived factor 1 (SDF-1) arechemotactic primarily for neutrophils and lymphocytes, whereasβ-chemokines, such as RANTES, MIP-1α, MIP-1β, monocyte chemotacticprotein-1 (MCP-1), MCP-2, MCP-3 and eotaxin are chemotactic formacrophages, T-cells, eosinophils and basophils (Deng, et al., Nature,381:661-666 (1996)). The C chemokine lymphotactin shows specificity forlymphocytes (Kelner, et al., Science, 266:1395-1399 (1994)) while theCX₃C chemokine fractalkine shows specificity for lymphocytes andmonocytes (Bazan, et al., Nature, 385:640-644 (1997).

Chemokines bind specific cell-surface receptors belonging to the familyof G-protein-coupled seven-transmembrane-domain proteins (reviewed inHoruk, Trends Pharm. Sci., 15:159-165 (1994)) termed “chemokinereceptors.” On binding their cognate ligands, chemokine receptorstransduce an intracellular signal through the associated heterotrimericG protein, resulting in a rapid increase in intracellular calciumconcentration. There are at least twelve human chemokine receptors thatbind or respond to β-chemokines with the following characteristicpattern: CCR1 (or “CKR-1” or “CC-CKR-1”) MIP-1α, MIP-1β, MCP-3, RANTES(Ben-Barruch, et al., J. Biol. Chem., 270:22123-22128 (1995); Neote, etal., Cell, 72:415425 (1993)); CCR2A and CCR2B (or “CKR-2A”/“CKR-2A” or“CC-CKR-2A”/“CC-CKR2A”) MCP-1, MCP-3, MCP-4; CCR3 (or “CKR-3” or“CC-CKR-3”) eotaxin, RANTES, MCP; (Ponath, et al., J. Exp. Med.,183:2437-2448 (1996)); CCR4 (or “CKR-4” or “CC-CKR-4”) TARC, MDC (Imai,et al., J. Biol. Chem., 273:1764-1768 (1998)); CCR5 (or “CKR-5” or“CC-CKR-5”) MIP-1α, RANTES, MIP-1β (Sanson, et al., Biochemistry,35:3362-3367 (1996)); CCR6 MIP-3 alpha (Greaves, et al., J. Exp. Med.,186:837-844 (1997)); CCR7 MIP-3 beta and 6Ckine (Campbell, et al., J.Cell. Biol., 141:1053-1059(1998)); CCR8 I-309, HHV8 vMIP-I, HHV-8vMIP-II, MCV vMCC-I (Dairaghi, et al., J. Biol. Chem., 274:21569-21574(1999)); CCR9 TECK (Zaballos, et al., J. Immunol., 162:5671-5675(1999)), D6 MIP-1 beta, RANTES, and MCP-3 (Nibbs, et al., J. Biol.Chem., 272:32078-32083 (1997)), and the Duffy blood-group antigenRANTES, MCP-1 (Chaudhun, et al., J. Biol. Chem., 269:7835-7838 (1994)).

Chemokine receptors, such as CCR1, CCR2, CCR2A, CCR2B, CCR3, CCR4, CCR5,CCR6, CCR7, CCR8, CCR9, CXCR1, CXCR2, CXCR3, CXCR4, CXCR5, CX₃CR1, andXCR1 have been implicated as being important mediators of inflammatoryand immunoregulatory disorders and diseases, including asthma andallergic diseases, as well as autoimmune pathologies such as rheumatoidarthritis and atherosclerosis.

The CXCR3 chemokine receptor is expressed primarily in T lymphocytes,and its functional activity can be measured by cytosolic calciumelevation or chemotaxis. The receptor was previously referred to as GPR9or CKR-L2. Its chromosomal location is unusual among the chemokinereceptors in being localized to Xq13. Ligands that have been identifiedthat are selective and of high affinity are the CXC chemokines, IP10,MIG and ITAC.

The highly selective expression of CXCR3 makes it an ideal target forintervention to interrupt inappropriate T cell trafficking. The clinicalindications for such intervention are in T-cell mediated autoimmunediseases such as multiple sclerosis, rheumatoid arthritis, and type Idiabetes. Inappropriate T-cell infiltration also occurs in psoriasis andother pathogenic skin inflammation conditions, although the diseases maynot be true autoimmune disorders. In this regard, up-regulation of IP-10expression in keratinocytes is a common feature in cutaneousimmunopathologies. Inhibition of CXCR3 can be beneficial in reducingrejection in organ transplantation. Ectopic expression of CXCR3 incertain tumors, especially subsets of B cell malignancies indicate thatselective inhibitors of CXCR3 will have value in tumor immunotherapy,particularly attenuation of metastasis.

In view of the clinical importance of CXCR3, the identification ofcompounds that modulate CXCR3 function represents an attractive avenueinto the development of new therapeutic agents. Such compounds areprovided herein.

SUMMARY OF THE INVENTION

The present invention provides compounds which are useful in thetreatment or prevention of certain inflammatory and immunoregulatorydisorders and diseases, including asthma and allergic diseases, as wellas autoimmune pathologies such as rheumatoid arthritis andatherosclerosis. The compounds provided herein have the general formula(I):

wherein X represents a bond, —C(O)—, —C(R⁵)(R⁶)—, —C(R⁵)═, —S(O)—,—S(O)₂— or —N═; Z represents a bond, —N═, —O—, —S—, —N(R¹⁷)— or —C(R⁷)═,with the proviso that X and Z are not both a bond; L represents a bond,C(O)—(C₁-C₈)alkylene, (C₁-C₈)alkylene or (C₂-C₈)heteroalkylene; Qrepresents a bond, (C₁-C₈)alkylene, (C₂-C₈)heteroalkylene, —C(O)—,—OC(O)—, —N(R⁸)C(O)—, —CH₂CO—, —CH₂SO— or —CH₂SO₂—, and optionally L andQ can be linked together to form a 5- or 6-membered heterocyclic grouphaving from 1 to 3 heteroatoms. The symbols R¹ and R² independentlyrepresent H, (C₁-C₈)alkyl, (C₂-C₈)heteroalkyl, aryl or heteroaryl, oroptionally are combined to form a 3 to 8-membered ring having from 0 to2 heteroatoms as ring vertices, and optionally R² and L can be linkedtogether to form a 5- or 6-membered heterocyclic group having from 1 to4 heteroatoms. The symbol R³ represents hydroxy, (C₁-C₈)alkoxy, amino,(C₁-C₈)alkylamino, di(C₁-C₈)alkylamino, (C₂-C₈)heteroalkyl,(C₃-C₉)heterocyclyl, (C₁-C₈)acylamino, amidino, guanidino, ureido,cyano, heteroaryl, —CONR⁹R¹⁰ or —CO₂R¹¹. The symbol R⁴ represents(C₁-C₂₀)alkyl, (C₂-C₂₀)heteroalkyl, heteroaryl, aryl,heteroaryl(C₁-C₆)alkyl, heteroaryl(C₂-C₆)heteroalkyl, aryl(C₁-C₆)alkylor aryl(C₂-C₆)heteroalkyl. The symbols R⁵ and R⁶ independently representH, (C₁-C₈)alkyl, (C₂-C₈)heteroalkyl, heteroaryl or aryl, or optionallyR⁵ and R⁶ are combined to form a 3- to 7-membered ring. The symbols R⁷and R⁸ independently represent H, (C₁-C₈)alkyl, (C₂-C₈)heteroalkyl,heteroaryl or aryl. The symbols R⁹, R¹⁰ and R¹¹ each independentlyrepresent H, (C₁-C₈)alkyl, (C₂-C₈)heteroalkyl, heteroaryl, aryl,heteroaryl(C₁-C₆)alkyl, heteroaryl(C₂-C₈)heteroalkyl, aryl(C₁-C₈)alkylor aryl(C₂-C₈)heteroalkyl.

Turning next to the ring vertices, Y¹, Y², Y³ and Y⁴, the symbols Y¹ andY² independently represent —C(R¹²)═, —N═, —O—, —S—, or —N(R¹³)—. Thesymbol Y³ represents N or C wherein the carbon atom shares a double bondwith either Z or Y⁴; and Y⁴ represents —N(R¹⁴)—, —C(R¹⁴)═, —N═ orN(R¹⁴)C(R¹⁵)(R¹⁶)—. In the above groups, the symbol R¹² represents H,halogen, hydroxy, amino, alkylamino, dialkylamino, (C₁-C₈)alkyl,(C₂-C₈)heteroalkyl, heteroaryl and aryl, or optionally when Y¹ and Y²are both —C(R¹²)═ the two R¹² groups can be combined to form asubstituted or unsubstituted 5- to 6-membered cycloalkyl,heterocycloalkyl, aryl or heteroaryl ring; or optionally when Y¹ is—C(R¹²)═ and X is —C(R⁵)═ or —C(R⁵)(R⁶)—, R¹² and R⁵ can be combined toform a substituted or unsubstituted 5- to 6-membered cycloalkyl,heterocycloalkyl, aryl or heteroaryl ring. Additionally, the symbol R¹³represents H, (C₁-C₈)alkyl, (C₂-C₈)heteroalkyl, heteroaryl, aryl,heteroaryl(C₁-C₆)alkyl, heteroaryl(C₂-C₈)heteroalkyl, aryl(C₁-C₈)alkylor aryl(C₂-C₈)heteroalkyl. The symbol R¹⁴ represents (C₁-C₈)alkyl,(C₂-C₈)heteroalkyl, aryl(C₁-C₈)alkyl, aryl(C₂-C₈)heteroalkyl,heteroaryl(C₁-C₈)alkyl, heteroaryl(C₂-C₈)heteroalkyl, heteroaryl andaryl; R¹⁵ and R¹⁶ are independently selected from H, (C₁-C₈)alkyl and(C₂-C₈)heteroalkyl; and R¹⁷ is selected from H, (C₁-C₈)alkyl,(C₂-C₈)heteroalkyl, heteroaryl, aryl, heteroaryl(C₁-C₆)alkyl,heteroaryl(C₂-C₈)heteroalkyl, aryl(C₁-C₈)alkyl andaryl(C₂-C₈)heteroalkyl, or optionally when Y² is —C(R¹²)═ or —N(R¹³)—,R¹⁷ can be combined with R¹² or R¹³ to form a substituted orunsubstituted 5- to 6-membered cycloalkyl, heterocycloalkyl, aryl orheteroaryl ring; with the proviso that when the Y³-containing ringsystem is a quinazolinone or quinolinone ring system, and R⁴—Q— issubstituted or unsubstituted (C₅-C₁₅)alkyl, then R³—L— is other thansubstituted or unsubstituted (C₂-C₈)alkylene or a substituted orunsubstituted (C₂-C₈)heteroalkylene attached to —NR′R″, wherein R′ andR″ are independently selected from the group consisting of hydrogen and(C₁-C₈)alkyl, or optionally are combined with the nitrogen atom to whicheach is attached to form a 5-, 6- or 7-membered ring.

Unless otherwise indicated, the compounds provided in the above formulaare meant to include pharmaceutically acceptable salts and prodrugsthereof.

The present invention also provides pharmaceutical compositionscomprising a compound of formula I and a pharmaceutically acceptableexcipient or carrier.

The present invention further provides methods for the treatment orprevention of an inflammatory or immune condition or disorder,comprising administering to a subject in need of such treatment orprevention a therapeutically effective amount of a compound of formulaI.

The present invention also provides methods for the treatment orprevention of a condition or disorder mediated by the CXCR3 chemokinereceptor, comprising administering to a subject in need of suchtreatment or prevention a therapeutically effective amount of a compoundof formula I.

The present invention also provides methods for the modulation of CXCR3,comprising contacting a cell with a compound of formula I.

The present invention further provides methods for the modulation ofCXCR3, comprising contacting a CXCR3 protein with a compound of formulaI.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 illustrates a general synthesis scheme for racemic substitutedquinazolinones of the invention.

FIG. 2 illustrates the generic synthesis of substituted quinolines ofthe invention.

FIG. 3 illustrates the generic synthesis of substituted naphthalenes ofthe invention.

FIG. 4 illustrates the generic synthesis of enantiomerically enrichedsubstituted quinazolinones and 8-aza-quinazolinones of the invention.

FIG. 5 illustrates the generic synthesis of substituted benzimidazolesof the invention.

FIG. 6 illustrates the synthesis of two regioisomeric substitutedthiazoles of the invention.

FIG. 7 illustrates the generic synthesis of substituted benzothiophenesof the invention.

FIG. 8 illustrates the generic synthesis of substituted imidazoles ofthe invention.

FIG. 9 illustrates the generic synthesis of substituted triazolinones ofthe invention.

FIG. 10 illustrates the generic synthesis of substituted purine-6-onesof the invention.

FIG. 11 illustrates the generic synthesis of regioisomeric (see FIG. 1)substituted quinazolinones of the invention.

FIG. 12 illustrates exemplary structures for certain compounds of theinvention.

FIG. 13 illustrates a representative synthesis of 8-azaquinazolinones ofthe invention.

FIGS. 14-18 illustrate synthetic routes for exemplary compounds of theinvention.

FIG. 19 provides a table showing the CXCR3 antagonist activity forexemplary compounds of the invention.

DETAILED DESCRIPTION OF THE INVENTION

Definitions

The term “alkyl,” by itself or as part of another substituent, means,unless otherwise stated, a straight or branched chain, or cyclichydrocarbon radical, or combination thereof, which may be fullysaturated, mono- or polyunsaturated and can include di- and multivalentradicals, having the number of carbon atoms designated (i.e. C₁-C₁₀means one to ten carbons). Examples of saturated hydrocarbon radicalsinclude groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl,t-butyl, isobutyl, sec-butyl, cyclohexyl, (cyclohexyl)methyl,cyclopropylmethyl, homologs and isomers of, for example, n-pentyl,n-hexyl, n-heptyl, n-octyl, and the like. An unsaturated alkyl group isone having one or more double bonds or triple bonds. Examples ofunsaturated alkyl groups include vinyl, 2-propenyl, crotyl,2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(1,4-pentadienyl),ethynyl, 1- and 3-propynyl, 3-butynyl, and the higher homologs andisomers.

The term “alkylene” by itself or as part of another substituent means adivalent radical derived from an alkane, as exemplified by—CH₂CH₂CH₂CH₂—, and further includes those groups described below as“heteroalkylene.” Typically, an alkyl (or alkylene) group will have from1 to 24 carbon atoms, with those groups having 10 or fewer carbon atomsbeing preferred in the present invention. A “lower alkyl” or “loweralkylene” is a shorter chain alkyl or alkylene group, generally havingeight or fewer carbon atoms, or more.

The terms “alkoxy,” “alkylamino” and “alkylthio” (or thioalkoxy) areused in their conventional sense, and refer to those alkyl groupsattached to the remainder of the molecule via an oxygen atom, an aminogroup, or a sulfur atom, respectively. Similarly, the term dialkylaminorefers to an amino group having two attached alkyl groups that can bethe same or different.

The term “heteroalkyl,” by itself or in combination with another term,means, unless otherwise stated, a stable straight or branched chain, orcyclic hydrocarbon radical, or combinations thereof, consisting of thestated number of carbon atoms and from one to three heteroatoms selectedfrom the group consisting of O, N, Si and S, and wherein the nitrogenand sulfur atoms may optionally be oxidized and the nitrogen heteroatommay optionally be quaternized. The heteroatom(s) O, N and S may beplaced at any interior position of the heteroalkyl group. The heteroatomSi may be placed at any position of the heteroalkyl group, including theposition at which the alkyl group is attached to the remainder of themolecule. Examples include —CH₂—CH₂—O—CH₃, —CH₂—CH₂—NH—CH₃,—CH₂—CH₂—N(CH₃)—CH₃, —CH₂—S—CH₂—CH₃, —CH₂—CH₂, —S(O)—CH₂—CH₂—S(O)₂—CH₃,—CH═CH—O—CH₃, —Si(CH₃)₃, —CH₂—CH═N—OCH₃, and —CH═CH—N(CH₃)—CH₃. Up totwo heteroatoms may be consecutive, such as, for example, —CH₂—NH—OCH₃and —CH₂—O—Si(CH₃)₃. When a prefix such as (C₂-C₈) is used to refer to aheteroalkyl group, the number of carbons (2-8, in this example) is meantto include the heteroatoms as well. For example, a C₂-heteroalkyl groupis meant to include, for example, —CH₂OH (one carbon atom and oneheteroatom replacing a carbon atom) and —CH₂SH. The term“heteroalkylene” by itself or as part of another substituent means adivalent radical derived from heteroalkyl, as exemplified by—CH₂—CH₂—S—CH₂CH₂— and —CH₂—S—CH₂—CH₂—NH—CH₂—. For heteroalkylenegroups, heteroatoms can also occupy either or both of the chain termini(e.g., alkyleneoxy, alkylenedioxy, alkyleneamino, alkylenediamino, andthe like). Still further, for alkylene and heteroalkylene linkinggroups, no orientation of the linking group is implied.

The terms “cycloalkyl” and “heterocycloalkyl”, by themselves or incombination with other terms, represent, unless otherwise stated, cyclicversions of “alkyl” and “heteroalkyl”, respectively. Additionally, forheterocycloalkyl, a heteroatom can occupy the position at which theheterocycle is attached to the remainder of the molecule. Examples ofcycloalkyl include cyclopentyl, cyclohexyl, 1-cyclohexenyl,3-cyclohexenyl, cycloheptyl, and the like. Examples of heterocycloalkylinclude 1-(1,2,5,6-tetrahydropyridyl), 1-piperidinyl, 2-piperidinyl,3-piperidinyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl,tetrahydrofuran-3-yl, tetrahydrothien-2-yl, tetrahydrothien-3-yl,1-piperazinyl, 2-piperazinyl, and the like.

The terms “halo” or “halogen,” by themselves or as part of anothersubstituent, mean, unless otherwise stated, a fluorine, chlorine,bromine, or iodine atom. Additionally, terms such as “haloalkyl,” aremeant to include monohaloalkyl and polyhaloalkyl. For example, the term“halo(C₁-C₄)alkyl” is mean to include trifluoromethyl,2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.

The term “aryl” means, unless otherwise stated, a polyunsaturated,typically aromatic, hydrocarbon substituent which can be a single ringor multiple rings (up to three rings) which are fused together or linkedcovalently. The term “heteroaryl” refers to aryl groups (or rings) thatcontain from zero to four heteroatoms selected from N, O, and S, whereinthe nitrogen and sulfur atoms are optionally oxidized, and the nitrogenatom(s) are optionally quaternized. A heteroaryl group can be attachedto the remainder of the molecule through a heteroatom. Non-limitingexamples of aryl and heteroaryl groups include phenyl, 1-naphthyl,2-naphthyl, 4-biphenyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 3-pyrazolyl,2-imidazolyl, 4-imidazolyl, pyrazinyl, 2-oxazolyl, 4-oxazolyl,2-phenyl-4-oxazolyl, 5-oxazolyl, 3-isoxazolyl, 4-isoxazolyl,5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-furyl, 3-furyl,2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl,4-pyrimidyl, 5-benzothiazolyl, purinyl, 2-benzimidazolyl, 5-indolyl,1-isoquinolyl, 5-isoquinolyl, 2-quinoxalinyl, 5-quinoxalinyl,3-quinolyl, and 6-quinolyl. Substituents for each of the above notedaryl and heteroaryl ring systems are selected from the group ofacceptable substituents described below.

For brevity, the term “aryl” when used in combination with other terms(e.g., aryloxy, arylthioxy, arylalkyl) includes both aryl and heteroarylrings as defined above. Thus, the term “arylalkyl” is meant to includethose radicals in which an aryl group is attached to an alkyl group(e.g., benzyl, phenethyl, pyridylmethyl and the like) including thosealkyl groups in which a carbon atom (e.g., a methylene group) has beenreplaced by, for example, an oxygen atom (e.g., phenoxymethyl,2-pyridyloxymethyl, 3-(1-naphthyloxy)propyl, and the like).

Each of the above terms (e.g., “alkyl,” “heteroalkyl,” “aryl” and“heteroaryl”) are meant to include both substituted and unsubstitutedforms of the indicated radical. Preferred substituents for each type ofradical are provided below.

Substituents for the alkyl and heteroalkyl radicals (including thosegroups often referred to as alkylene, alkenyl, heteroalkylene,heteroalkenyl, alkynyl, cycloalkyl, heterocycloalkyl, cycloalkenyl, andheterocycloalkenyl) can be a variety of groups selected from: —OR′, ═O,═NR′, ═N—OR′, —NR′R″, —SR′, -halogen, -SiR′R″R′″, —OC(O)R′, —C(O)R′,—CO₂R′, —CONR′R″, —OC(O)NR′R″, —NR″C(O)R′, —NR′—C(O)NR″R′″, —NR″C(O)₂R′,—NH—C(NH₂)═NH, —NR′C(NH₂)═NH, —NH—C(NH₂)═NR′, —S(O)R′, —S(O)₂R′,—S(O)₂NR′R″, —CN and —NO₂ in a number ranging from zero to (2 m′+1),where m′ is the total number of carbon atoms in such radical. R′, R″ andR′″each independently refer to H, unsubstituted (C₁-C₈)alkyl andheteroalkyl, unsubstituted aryl, aryl substituted with 1-3 halogens,alkoxy or thioalkoxy groups, or aryl-(C₁-C₄)alkyl groups. When R′ and R″are attached to the same nitrogen atom, they can be combined with thenitrogen atom to form a 5-, 6-, or 7-membered ring. For example,—NR′R″is meant to include 1-pyrrolidinyl and 4-morpholinyl. From theabove discussion of substituents, one of skill in the art willunderstand that the term “alkyl” in its broadest sense is meant toinclude groups such as haloalkyl (e.g., —CF₃ and —CH₂CF₃) and acyl(e.g., —C(O)CH₃, —C(O)CF₃, —C(O)CH₂OCH₃, and the like). Preferably, thealkyl groups will have from 0-3 substituents, more preferably 0, 1, or 2substituents, unless otherwise specified.

Similarly, substituents for the aryl and heteroaryl groups are variedand are selected from: -halogen, —OR′, —OC(O)R′, —NR′R″, —SR′, —R′, —CN,—NO₂, —CO₂R′, —CONR′R″, —C(O)R′, —OC(O)NR′R″, —NR″C(O)R′, —NR″C(O)₂R′,—NR′—C(O)NR″R′″, —NH—C(NH₂)═NH, —NR′C(NH₂)═NH, —NH—C(NH₂)═NR′, —S(O)R′,—S(O)₂R′, —S(O)₂NR′R″, —N₃, —CH(Ph)₂, perfluoro(C₁-C₄)alkoxy, andperfluoro(C₁-C₄)alkyl, in a number ranging from zero to the total numberof open valences on the aromatic ring system; and where R′, R″ and R′″are independently selected from H, (C₁-C₈)alkyl and heteroalkyl,unsubstituted aryl and heteroaryl, (unsubstituted aryl)-(C₁-C₄)alkyl,and (unsubstituted aryl)oxy-(C₁-C₄)alkyl.

Two of the substituents on adjacent atoms of the aryl or heteroaryl ringmay optionally be replaced with a substituent of the formula—T—C(O)—(CH₂)_(q)—U—, wherein T and U are independently —NH—, —O—, —CH₂—or a single bond, and q is an integer of from 0 to 2. Alternatively, twoof the substituents on adjacent atoms of the aryl or heteroaryl ring mayoptionally be replaced with a substituent of the formula—A—(CH₂)_(r)—B—, wherein A and B are independently —CH₂—, —O—, —NH—,—S—, —S(O)—, —S(O)₂—, —S(O)₂NR′— or a single bond, and r is an integerof from 1 to 3. One of the single bonds of the new ring so formed mayoptionally be replaced with a double bond. Alternatively, two of thesubstituents on adjacent atoms of the aryl or heteroaryl ring mayoptionally be replaced with a substituent of the formula—(CH₂)_(s)—X—(CH₂)_(t)—, where s and t are independently integers offrom 0 to 3, and X is —O—, —NR′—, —S—, —S(O)—, —S(O)₂—, or —S(O)₂NR′—.The substituent R′ in —NR′— and —S(O)₂NR′— is selected from hydrogen orunsubstituted (C₁-C₆)alkyl.

As used herein, the term “heteroatom” is meant to include oxygen (O),nitrogen (N), sulfur (S) and silicon (Si).

The term “pharmaceutically acceptable salts” is meant to include saltsof the active compounds which are prepared with relatively nontoxicacids or bases, depending on the particular substituents found on thecompounds described herein. When compounds of the present inventioncontain relatively acidic functionalities, base addition salts can beobtained by contacting the neutral form of such compounds with asufficient amount of the desired base, either neat or in a suitableinert solvent. Examples of pharmaceutically acceptable base additionsalts include sodium, potassium, calcium, ammonium, organic amino, ormagnesium salt, or a similar salt. When compounds of the presentinvention contain relatively basic functionalities, acid addition saltscan be obtained by contacting the neutral form of such compounds with asufficient amount of the desired acid, either neat or in a suitableinert solvent. Examples of pharmaceutically acceptable acid additionsalts include those derived from inorganic acids like hydrochloric,hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric,monohydrogenphosphoric, dihydrogenphosphoric, sulfuric,monohydrogensulfuric, hydriodic, or phosphorous acids and the like, aswell as the salts derived from relatively nontoxic organic acids likeacetic, propionic, isobutyric, maleic, malonic, benzoic, succinic,suberic, fumaric, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic,citric, tartaric, methanesulfonic, and the like. Also included are saltsof amino acids such as arginate and the like, and salts of organic acidslike glucuronic or galactunoric acids and the like (see, for example,Berge, et al. (1977) J. Pharm. Sci. 66:1-19). Certain specific compoundsof the present invention contain both basic and acidic functionalitiesthat allow the compounds to be converted into either base or acidaddition salts.

(1) The terms “treat”, “treating” or “treatment”, as used herein, referto a method of alleviating or abrogating a disease and/or its attendantsymptoms. The terms “prevent”, “preventing” or “prevention”, as usedherein, refer to a method of barring a subject from acquiring a disease.

The neutral forms of the compounds may be regenerated by contacting thesalt with a base or acid and isolating the parent compound in theconventional manner. The parent form of the compound differs from thevarious salt forms in certain physical properties, such as solubility inpolar solvents, but otherwise the salts are equivalent to the parentform of the compound for the purposes of the present invention.

In addition to salt forms, the present invention provides compoundswhich are in a prodrug form. Prodrugs of the compounds described hereinare those compounds that readily undergo chemical changes underphysiological conditions to provide the compounds of the presentinvention. Additionally, prodrugs can be converted to the compounds ofthe present invention by chemical or biochemical methods in an ex vivoenvironment. For example, prodrugs can be slowly converted to thecompounds of the present invention when placed in a transdermal patchreservoir with a suitable enzyme or chemical reagent. Prodrugs are oftenuseful because, in some situations, they may be easier to administerthan the parent drug. They may, for instance, be bioavailable by oraladministration whereas the parent drug is not. The prodrug may also haveimproved solubility in pharmacological compositions over the parentdrug. A wide variety of prodrug derivatives are known in the art, suchas those that rely on hydrolytic cleavage or oxidative activation of theprodrug. An example, without limitation, of a prodrug would be acompound of the present invention which is administered as an ester (the“prodrug”), but then is metabolically hydrolyzed to the carboxylic acid,the active entity. Additional examples include peptidyl derivatives of acompound of the invention.

Certain compounds of the present invention can exist in unsolvated formsas well as solvated forms, including hydrated forms. In general, thesolvated forms are equivalent to unsolvated forms and are intended to beencompassed within the scope of the present invention. Certain compoundsof the present invention may exist in multiple crystalline or amorphousforms. In general, all physical forms are equivalent for the usescontemplated by the present invention and are intended to be within thescope of the present invention.

Certain compounds of the present invention possess asymmetric carbonatoms (optical centers) or double bonds; the racemates, diastereomers,geometric isomers and individual isomers are all intended to beencompassed within the scope of the present invention.

The compounds of the present invention may also contain unnaturalproportions of atomic isotopes at one or more of the atoms thatconstitute such compounds. For example, the compounds may beradiolabeled with radioactive isotopes, such as for example tritium(³H), iodine-125 (¹²⁵I) or carbon-14 (¹⁴C). Radiolabled compounds areuseful as therapeutic agents, e.g., cancer therapeutic agents, researchreagents, e.g., binding assay reagents, and diagnostic agents, e.g., invivo imaging agents. All isotopic variations of the compounds of thepresent invention, whether radioactive or not, are intended to beencompassed within the scope of the present invention.

Embodiments of the Invention

The present invention is directed to compounds, compositions and methodsuseful in the modulation of chemokine receptor activity, particularlyCXCR3. Accordingly, the compounds of the present invention are thosewhich inhibit at least one function or characteristic of a mammalianCXCR3 protein, for example, a human CXCR3 protein. The ability of acompound to inhibit such a function can be demonstrated in a bindingassay (e.g., ligand binding or agonist binding), a signalling assay(e.g., activation of a mammalian G protein, induction of rapid andtransient increase in the concentration of cytosolic free calcium),and/or cellular response function (e.g., stimulation of chemotaxis,exocytosis or inflammatory mediator release by leukocytes).

Compounds

The present invention provides compounds that are useful as antagonistsof CXCR3, having particular utility for the treatment or prevention ofinflammation. The compounds provided herein have the general formula(I):

wherein X represents a bond, —C(O)—, —C(R⁵)(R⁶)—, —C(R⁵)═, —S(O)—,—S(O)₂— or —N═; Z represents a bond, —N═, —O—, —S—, —N(R¹⁷)— or —C(R⁷)═,with the proviso that X and Z are not both a bond; L represents a bond,C(O)—(C₁-C₈)alkylene, (C₁-C₈)alkylene or (C₂-C₈)heteroalkylene; Qrepresents a bond, (C₁-C₈)alkylene, (C₂-C₈)heteroalkylene, —C(O)—,—OC(O)—, —N(⁸)C(O)—, —CH₂CO—, —CH₂SO—, or —CH₂SO₂—; and optionally L andQ can be linked together to form a 5- or 6-membered heterocyclic grouphaving from 1 to 3 heteroatoms. The symbols R¹ and R² independentlyrepresent H, (C₁-C₈)alkyl, (C₂-C₈)heteroalkyl, aryl or heteroaryl, oroptionally are combined to form a 3 to 8-membered ring having from 0 to2 heteroatoms as ring vertices, and optionally R² can be linked togetherwith L to form a 5- or 6-membered heterocyclic group having from 1 to 4heteroatoms. The symbol R³ represents hydroxy, (C₁-C₈)alkoxy, amino,(C₁-C₈)alkylamino, di(C₁-C₈)alkylamino, (C₂-C₈)heteroalkyl,(C₃-C₉)heterocyclyl, (C₁-C₈)acylamino, amidino, guanidino, ureido,cyano, heteroaryl, —CONR⁹R¹⁰ or —CO₂R¹¹. The symbol R⁴ represents(C₁-C₂₀)alkyl, (C₂-C₂₀)heteroalkyl, heteroaryl, aryl,heteroaryl(C₁-C₆)alkyl, heteroaryl(C₂-C₆)heteroalkyl, aryl(C₁-C₆)alkylor aryl(C₂-C₆)heteroalkyl. The symbols R⁵ and R⁶ independently representH, (C₁-C₈)alkyl, (C₂-C₈)heteroalkyl, heteroaryl or aryl, or optionallyR⁵ and R⁶ are combined to form a 3- to 7-membered ring. The symbols R⁷and R⁸ independently represent H, (C₁-C₈)alkyl, (C₂-C₈)heteroalkyl,heteroaryl or aryl. The symbols R⁹, R¹⁰ and R¹¹ each independentlyrepresent H, (C₁-C₈)alkyl, (C₂-C₈)heteroalkyl, heteroaryl, aryl,heteroaryl(C₁-C₆)alkyl, heteroaryl(C₂-C₈)heteroalkyl, aryl(C₁-C₈)alkylor aryl(C₂-C₈)heteroalkyl.

Turning next to the ring vertices, Y¹, Y², Y³ and Y⁴, the symbols Y¹ andY² independently represent —C(R¹²)═, —N═, —O—, —S—, or —N(R¹³)—. Thesymbol Y³ represents N or C wherein the carbon atom shares a double bondwith either Z or Y⁴; and Y⁴ represents —N(R¹⁴)—, —C(R¹⁴)═, —N═ or—N(R¹⁴)—C(R¹⁵)(R¹⁶)—. In the above groups, the symbol R¹² represents H,halogen, hydroxy, amino, allylamino, dialkylamino, (C₁-C₈)alkyl,(C₂-C₈)heteroalkyl, heteroaryl and aryl, or optionally when Y¹ and Y²are both —C(R¹²)═ the two R¹² groups can be combined to form asubstituted or unsubstituted 5- to 6-membered cycloalkyl,heterocycloalkyl, aryl or heteroaryl ring; or optionally when Y¹ is—C(R¹²)═ and X is —C(R⁵)═ or —C(R⁵)(R⁶)—, R¹² and R⁵ can be combined toform a substituted or unsubstituted 5- to 6-membered cycloalkyl,heterocycloalkyl, aryl or heteroaryl ring. Additionally, the symbol R¹³represents H, (C₁-C₈)alkyl, (C₂-C₈)heteroalkyl, heteroaryl, aryl,heteroaryl(C₁-C₆)alkyl, heteroaryl(C₂-C₈)heteroalkyl, aryl(C₁-C₈)alkylor aryl(C₂-C₈)heteroalkyl. The symbol R¹⁴ represents (C₁-C₈)alkyl,(C₂-C₈)heteroalkyl, aryl(C₁-C₈)alkyl, aryl(C₂-C₈)heteroalkyl,heteroaryl(C₁-C₈)alkyl, heteroaryl(C₂-C₈)heteroalkyl, heteroaryl andaryl; R¹⁵ and R¹⁶ are independently selected from H, (C₁-C₈)alkyl and(C₂-C₈)heteroalkyl; and R¹⁷ is selected from H, (C₁-C₈)alkyl,(C₂-C₈)heteroalkyl, heteroaryl, aryl, heteroaryl(C₁-C₆)alkyl,heteroaryl(C₂-C₈)heteroalkyl, aryl(C₁-C₈)alkyl andaryl(C₂-C₈)heteroalkyl, or optionally when Y² is —C(R¹²)═ or —N(R¹³)—,R¹⁷ can be combined with R¹² or R¹³ to form a substituted orunsubstituted 5- to 6-membered cycloalkyl, heterocycloalkyl, aryl orheteroaryl ring; with the proviso that when the Y³-containing ringsystem is a quinazolinone or quinolinone ring system, and R⁴—Q— issubstituted or unsubstituted (C₅-C₁₅)alkyl, then R³—L— is other thansubstituted or unsubstituted (C₂-C₈)alkylene or a substituted orunsubstituted (C₂-C₈)heteroalkylene attached to —NR′R″, wherein R′ andR″ are independently selected from the group consisting of hydrogen and(C₁-C₈)alkyl, or optionally are combined with the nitrogen atom to whicheach is attached to form a 5-, 6- or 7-membered ring.

Embodiments represented by the above formula can be appreciated byreplacing the ring system having vertices X, Z, Y¹, Y², Y³ and Y⁴ withan appropriate scaffold wherein the attachment points represent theattachment of a R¹⁴ group and the carbon atom that bears the R¹ and R²groups:

For example, the ring system or “scaffold” is meant to include thefollowing (including substituted versions thereof) wherein the “A” ringis selected from those embodiments shown as:

Still other A ring scaffolds are six-membered rings (without additionalfused rings) and include:

In other embodiments, the A ring scaffolds are five-membered rings(without additional fused rings) and include, for example:

Typically, the ring substituents (shown as R and R′ groups in the abovefive-membered rings, but not shown in the fused ring sets orsix-membered rings above) are designed to provide electronic and/oradditional hydrophobic or hydrophilic character to the molecule to bringthe overall physical characters into conformity with those of the mostpreferred compounds in the series (see Examples).

Within each of the above groups of embodiments, R¹⁴ is preferably asubstituted or unsubstituted aryl group or a substituted orunsubstituted heteroaryl group. More preferably, the aryl or heteroarylgroups will have from 0 to 3 substituents. Still more preferably, 1 or 2substituents. The aryl and heteroaryl groups are preferably selectedfrom phenyl, substituted phenyl, pyridyl, substituted pyridyl,thiazolyl, substituted thiazolyl, pyrimidinyl, substituted pyrimidinyl,thienyl and substituted thienyl. For those embodiments having onesubstituent, the substituent will preferably be in a position para tothe point of attachment to the heterocyclic scaffolding. In the mostpreferred embodiments, the substituents are selected from cyano,halogen, (C₁-C₈)alkoxy, (C₁-C₈)alkyl, (C₂-C₈)heteroalkyl, CONH₂,methylenedioxy and ethylenedioxy.

Returning to formula I, in one group of preferred embodiments, X is—C(O)—. In another group, Z is —N═. In still another group of preferredembodiments, Y¹ and Y² are each —C(R¹²)═, wherein the two R¹² groups arecombined to form a fused 6-membered aryl or heteroaryl ring.Particularly preferred, are those embodiments that combine each of thesepreferred groups. Accordingly, in one group of particularly preferredembodiments, X is —C(O)—; Z is —N═; Y³ is C; and Y¹ and Y² are each—C(R¹²)═ wherein the two R¹² groups are combined to form a fused6-membered substituted or unsubstituted aryl or heteroaryl ring.

In other separate, but preferred embodiments, L is (C₁-C₈)alkylene; Q is—C(O)—, R⁴ is (C₅-C₁₅)alkyl, substituted or unsubstituted phenyl, orbiphenyl; R³ is (C₁-C₈)alkoxy, (C₁-C₈)alkylamino, di(C₁-C₈)alkylamino,(C₂-C₈)heteroalkyl, (C₃-C₉)heterocyclyl, (C₁-C₈)acylamino, cyano,heteroaryl, —CONR⁹R¹⁰ or —CO₂R¹¹; R¹ and R² are each independently H or(C₁-C₄)alkyl; Y³ is C and the carbon atom shares a double bond with Z;and the Y³-containing ring system is selected from quinoline,quinazoline, naphthalene, quinolinone, quinazolinone, triazolinone,pyrimidin-4-one, benzimidazole, thiazole, imidazole, pyridine, pyrazineand benzodiazepine.

Still other preferred embodiments can be defined according to the A ringscaffolding. For example, one group of preferred embodiments are thosein which X is —C(O)—; Z is —N═; Y³ is C; and Y¹ and Y² are each—C(R¹²)═. More preferably, the two R¹² groups are combined to form afused 6-membered substituted or unsubstituted aryl or heteroaryl ring.Particularly preferred are those embodiments in which Y⁴ is —N(R¹⁴)— or—C(R¹⁴)═ wherein the R¹⁴ group is a substituted or unsubstituted aryl orheteroaryl. In another group of preferred embodiments, X is —C(R⁵)(R⁶)—;Y⁴ is —N(R¹⁴)—, wherein R¹⁴ is substituted or unsubstituted aryl orheteroaryl; Y³ is C; Z is —N═; and Y¹ and Y² are each —C(R¹²)═. Inanother group of preferred embodiments, X is —C(R⁵)═; Y⁴ is —C(R¹⁴)═,wherein R¹⁴ is substituted or unsubstituted aryl or heteroaryl; Y³ is C;Z is —N═; and Y¹ and Y² are each —C(R¹²)═. In another group of preferredembodiments, X is a bond; Y⁴ is —N(R¹⁴)—, wherein R¹⁴ is substituted orunsubstituted aryl or heteroaryl; Y³ is C; Z is —N═; and Y¹ and Y² areeach —C(R¹²)═. In another group of preferred embodiments, X is —C(R⁵)═;Y⁴ is —C(R¹⁴)═, wherein R¹⁴ is substituted or unsubstituted aryl orheteroaryl; Y³ is C; Z is —C(R⁷)═; and Y¹ and Y² are each —C(R¹²)═. Inanother group of preferred embodiments, X is a bond; Z is —N═ or—N(R¹⁷)—; Y⁴ is —C(R¹⁴)═, wherein R¹⁴ is substituted or unsubstitutedaryl or heteroaryl; Y¹ is selected from the group consisting of —O—, —S—and —N(R¹³)— and Y² is —C(R¹²)—. In this group of embodiments, furtherpreferred are those compounds in which Y¹ is —O— and Z is —N═; compoundsin which Y¹ is —S— and Z is —N═; and compounds in which Y¹ is —N(R¹³)—and Z is —N═. In another group of preferred embodiments, X is —SO₂—; Y⁴is —N(R¹⁴)═, wherein R¹⁴ is substituted or unsubstituted aryl orheteroaryl; Y³ is C; Z is —N═ or —C(R⁷)═; and Y¹ and Y² are each—C(R¹²)═. In another group of preferred embodiments, X is a bond; Z is—O—, —S— or —N(R¹⁷)—; Y¹ is —N═ or —N(R¹³)—; Y² is —C(R¹²)═; and Y⁴ is—C(R¹⁴)═ wherein R¹⁴ is substituted or unsubstituted aryl or heteroaryl.Particularly preferred embodiments in this group are those in which Y¹is —N═ and Z is —O—; those in which Y¹ is —N═ and Z is —S—; and those inwhich Z is —N(R¹⁷)—. In another group of preferred embodiments, X is abond; Y¹ is —N(R¹³)— or ═N—; Y² is —C(R¹²)═; Y³ is C; Y⁴ is —C(R¹⁴)═wherein R¹⁴ is substituted or unsubstituted aryl or heteroaryl; and Z is—N(R¹⁷)— or ═N—, with the proviso that Y¹ and Z are not both ═N—. Inanother group of preferred embodiments, X is a bond; Y¹ and Y² are eachindependently —C(R¹²)═; Y³ is C; Y⁴ is —C(R¹⁴)— wherein R¹⁴ issubstituted or unsubstituted aryl or heteroaryl; and Z is —N(R¹⁷)—, O orS. More preferably, the two R¹² groups are combined to form a fused 5-or 6-membered substituted or unsubstituted aryl or heteroaryl ring. Inanother group of preferred embodiments, X is —C(O)—; Y¹ is —N(R¹³)—; Y²is —N═; Y³ is C; Y⁴ is —N(R¹⁴)— wherein R¹⁴ is substituted orunsubstituted aryl or heteroaryl; and Z is a bond. In another group ofpreferred embodiments, X is —C(O)—; Z is —N(R¹⁷)— wherein R¹⁷ issubstituted or unsubstituted aryl or heteroaryl; Y¹ and Y² are eachindependently —C(R¹²)═; Y³ is C; and Y⁴ is —N═. In another group ofpreferred embodiments, X and Z are —N═, Y¹ and Y² are each independently—C(R¹²)═; Y³ is C; and Y⁴ is —C(R¹⁴)═ wherein R¹⁴ is a substituted orunsubstituted aryl or heteroaryl group. In another group of preferredembodiments, wherein X is —C(O)—; Y⁴ is —N(R¹⁴)—C(R⁵)(R⁶)—; wherein R¹⁴is substituted or unsubstituted aryl or heteroaryl; Y¹ and Y² are eachindependently —C(R¹²)═; Y³ is C; and Z is —N═.

In each of the above groups of preferred embodiments, R¹ is mostpreferably H.

In one particularly preferred group of embodiments, the A ring is afused 6,6 or 6,5-member ring system having the indicated nitrogenvertices (see formula II).

In formula II, each of A¹, A², A³ and A⁴ is independently C or N.Preferably, no more than two of A¹-A⁴ are N. Additionally, X is —CO—,—CH₂— or a bond; R¹ and R² are each independently H or (C₁-C₄)alkyl; R¹⁴is a substituted or unsubstituted phenyl, pyridyl, thiazolyl, thienyl orpyrimidinyl group; Q is —CO—; L is (C₁-C₈)alkylene; the subscript n isan integer of from 0 to 4; and each R_(a) is independently selected fromhalogen, —OR′, —OC(O)R′, —NR′R″, —SR′, —R′, —CN, —NO₂, —CO₂R′, —CONR′R″,—C(O)R′, —OC(O)NR′R″, —NR″C(O)R′, —NR″C(O)₂R′, —NR″C(O)NR″R′″,—NH—C(NH₂)═NH, —NR′C(NH₂)═NH, —NHC(NH₂)═NR′, —S(O)R′, —S(O)₂R′,—S(O)₂NR′R″, —N₃, —CH(Ph)₂, perfluoro(C₁-C₄)alkoxy, andperfluoro(C₁-C₄)alkyl, wherein R′, R″ and R′″ are each independentlyselected from H, (C₁-C₈)alkyl, (C₂-C₈)heteroalkyl, unsubstituted aryl,unsubstituted heteroaryl, (unsubstituted aryl)-(C₁-C₄)alkyl, and(unsubstituted aryl)oxy-(C₁-C₄)alkyl. The remaining symbols, R³ and R⁴,have the meanings (and preferred groupings) provided above.

Still more preferably, the compound has the formula (III):

wherein A⁴ is C or N; X is —CO—, —CH₂— or a bond; R¹ and R² are eachindependently H or (C₁-C₄)alkyl; R¹⁴ is a substituted or unsubstitutedphenyl, pyridyl, thiazolyl, thienyl or pyrimidinyl group; Q is —CO—; Lis (C₁-C₈)alkylene; the subscript n is an integer of from 0 to 4; andeach R_(a) is independently selected from the group consisting ofhalogen, —OR′, —OC(O)R′, —NR′R″, —SR′, —R′, —CN, —NO₂, —CO₂R′, —CONR′R″,—C(O)R′, —OC(O)NR′R″, —NR″C(O)R′, —NR″C(O)₂R′, —NR′—C(O)NR″R′″,—NH—C(NH₂)═NH, —NR′C(NH₂)═NH, —NHC(NH₂)═NR′, —S(O)R′, —S(O)₂R′,—S(O)₂NR′R″, —N₃, —CH(Ph)₂, perfluoro(C₁-C₄)alkoxy, andperfluoro(C₁-C₄)alkyl, wherein R′, R″ and R′″ are each independentlyselected from H, (C₁-C₈)alkyl, (C₂-C₈)heteroalkyl, unsubstituted aryl,unsubstituted heteroaryl, (unsubstituted aryl)-(C₁-C₄)alkyl, and(unsubstituted aryl)oxy-(C₁-C₄)alkyl. The remaining symbols, R³ and R⁴,have the meanings (and preferred groupings) provided above.

In one group of preferred embodiments, X is —CO—. In another group ofpreferred embodiments, X is —CH₂—. In yet another group of preferredembodiments, X is a bond.

Further preferred compounds of formula III are those in which R¹ ismethyl, ethyl or propyl and R² is hydrogen or methyl. More preferably,R¹ and R² are each methyl. Still other preferred compounds of formulaIII are those in which R³ is selected from substituted or unsubstitutedpyridyl or substituted or unsubstituted imidazolyl. Also preferred arethose compounds of formula III in which R⁴ is a substituted orunsubstituted benzyl group, wherein the substituents are selected fromhalogen, halo(C₁-C₄)alkyl, halo(C₁-C₄)alkoxy, cyano, nitro, and phenyl.A preferred group for L is (C₁-C₄)alkylene. Also preferred are thosecompounds of formula III in which R¹⁴ is selected from substitutedphenyl, substituted pyridyl, substituted thiazolyl and substitutedthienyl, wherein the substituents are selected from cyano, halogen,(C₁-C₈)alkoxy, (C₁-C₈)alkyl, (C₂-C₈)heteroalkyl, CONH₂, methylenedioxyand ethylenedioxy. Still further preferred are those compounds thatcombine two or more of the preferred groups listed above.

In particularly preferred embodiments for compounds of formula III, X is—CO—; R¹ and R² are each independently selected from the groupconsisting of H, methyl and ethyl; R¹⁴ is selected from the groupconsisting of substituted or unsubstituted phenyl; Q is —CO—; L ismethylene, ethylene or propylene, R³ is selected from the groupconsisting of substituted or unsubstituted pyridyl and substituted orunsubstituted imidazolyl; R⁴ is substituted or unsubstituted benzyl,wherein said substituents are selected from the group consisting ofhalogen, halo(C₁-C₄)alkyl, halo(C₁-C₄)alkoxy, cyano, nitro, and phenyl;and each R_(a) is selected from the group consisting of halogen, —OR′,—OC(O)R′, —NR′R″, —SR′, —R′, —CN, —NO₂, —CO₂R′, —CONR′R″, —C(O)R′,—NR″C(O)R′, —NR′—C(O)NR″R′″, perfluoro(C₁-C₄)alkoxy, andperfluoro(C₁-C₄)alkyl, wherein R′, R″ and R′″ are each independentlyselected from the group consisting of H, (C₁-C₈)alkyl,(C₂-C₈)heteroalkyl, unsubstituted aryl, unsubstituted heteroaryl,(unsubstituted aryl)-(C₁-C₄)alkyl, and (unsubstitutedaryl)oxy-(C₁-C₄)alkyl.

Exemplary structures within this preferred group of embodiments are:

Preparation of the Compounds

FIGS. 1-18 provide a variety of synthesis routes to the compoundsprovided herein. One of skill in the art will appreciate that thesubstituents (e.g., R′, R″, R′″, R^(IV), etc.) can be altered before,during or after preparation of the heterocyclic scaffolding and thatsuitable adjustments in the exemplary conditions (e.g., temperatures,solvents, etc.) can be made. Additionally, one of skill in the art willrecognize that protecting groups may be necessary for the preparation ofcertain compounds and will be aware of those conditions compatible witha selected protecting group.

The exemplary methods and the examples described herein are illustrativeof the present invention and are not be construed as limiting the scopethereof.

Compositions

In another aspect, the present invention provides pharmaceuticalcompositions for modulating chemokine receptor activity in humans andanimals. The compositions comprise a compound of the present inventionwith a pharmaceutically acceptable carrier or diluent.

“Modulation” or modulating of chemokine receptor activity, as usedherein in its various forms, is intended to encompass antagonism,agonism, partial antagonism and/or partial agonism of the activityassociated with a particular chemokine receptor, preferably the CXCR3receptor. The term “composition” as used herein is intended to encompassa product comprising the specified ingredients (and in the specifiedamounts, if indicated), as well as any product which results, directlyor indirectly, from combination of the specified ingredients in thespecified amounts. By “pharmaceutically acceptable” it is meant thecarrier, diluent or excipient must be compatible with the otheringredients of the formulation and not deleterious to the recipientthereof.

The pharmaceutical compositions for the administration of the compoundsof this invention may conveniently be presented in unit dosage form andmay be prepared by any of the methods well known in the art of pharmacy.All methods include the step of bringing the active ingredient intoassociation with the carrier which constitutes one or more accessoryingredients. In general, the pharmaceutical compositions are prepared byuniformly and intimately bringing the active ingredient into associationwith a liquid carrier or a finely divided solid carrier or both, andthen, if necessary, shaping the product into the desired formulation. Inthe pharmaceutical composition the active object compound is included inan amount sufficient to produce the desired effect upon the process orcondition of diseases.

The pharmaceutical compositions containing the active ingredient may bein a form suitable for oral use, for example, as tablets, troches,lozenges, aqueous or oily suspensions, dispersible powders or granules,emulsions, hard or soft capsules, or syrups or elixirs. Compositionsintended for oral use may be prepared according to any method known tothe art for the manufacture of pharmaceutical compositions and suchcompositions may contain one or more agents selected from the groupconsisting of sweetening agents, flavoring agents, coloring agents andpreserving agents in order to provide pharmaceutically elegant andpalatable preparations. Tablets contain the active ingredient inadmixture with non-toxic pharmaceutically acceptable excipients whichare suitable for the manufacture of tablets. These excipients may be,for example, inert diluents, such as calcium carbonate, sodiumcarbonate, lactose, calcium phosphate or sodium phosphate; granulatingand disintegrating agents, for example, corn starch, or alginic acid;binding agents, for example starch, gelatin or acacia, and lubricatingagents, for example magnesium stearate, stearic acid or talc. Thetablets may be uncoated or they may be coated by known techniques todelay disintegration and absorption in the gastrointestinal tract andthereby provide a sustained action over a longer period. For example, atime delay material such as glyceryl monostearate or glyceryl distearatemay be employed. They may also be coated by the techniques described inU.S. Pat. Nos. 4,256,108; 4,166,452 and 4,265,874 to form osmotictherapeutic tablets for control release.

Formulations for oral use may also be presented as hard gelatin capsuleswherein the active ingredient is mixed with an inert solid diluent, forexample, calcium carbonate, calcium phosphate or kaolin, or as softgelatin capsules wherein the active ingredient is mixed with water or anoil medium, for example peanut oil, liquid paraffin, or olive oil.

Aqueous suspensions contain the active materials in admixture withexcipients suitable for the manufacture of aqueous suspensions. Suchexcipients are suspending agents, for example sodiumcarboxymethylcellulose, methylcellulose, hydroxy-propylmethylcellulose,sodium alginate, polyvinyl-pyrrolidone, gum tragacanth and gum acacia;dispersing or wetting agents may be a naturally-occurring phosphatide,for example lecithin, or condensation products of an alkylene oxide withfatty acids, for example polyoxy-ethylene stearate, or condensationproducts of ethylene oxide with long chain aliphatic alcohols, forexample heptadecaethyleneoxycetanol, or condensation products ofethylene oxide with partial esters derived from fatty acids and ahexitol such as polyoxyethylene sorbitol monooleate, or condensationproducts of ethylene oxide with partial esters derived from fatty acidsand hexitol anhydrides, for example polyethylene sorbitan monooleate.The aqueous suspensions may also contain one or more preservatives, forexample ethyl, or n-propyl, p-hydroxybenzoate, one or more coloringagents, one or more flavoring agents, and one or more sweetening agents,such as sucrose or saccharin.

Oily suspensions may be formulated by suspending the active ingredientin a vegetable oil, for example arachis oil, olive oil, sesame oil orcoconut oil, or in a mineral oil such as liquid paraffin. The oilysuspensions may contain a thickening agent, for example beeswax, hardparaffin or cetyl alcohol. Sweetening agents such as those set forthabove, and flavoring agents may be added to provide a palatable oralpreparation. These compositions may be preserved by the addition of ananti-oxidant such as ascorbic acid.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water provide the active ingredient inadmixture with a dispersing or wetting agent, suspending agent and oneor more preservatives. Suitable dispersing or wetting agents andsuspending agents are exemplified by those already mentioned above.Additional excipients, for example sweetening, flavoring and coloringagents, may also be present.

The pharmaceutical compositions of the invention may also be in the formof oil-in-water emulsions. The oily phase may be a vegetable oil, forexample olive oil or arachis oil, or a mineral oil, for example liquidparaffin or mixtures of these. Suitable emulsifying agents may benaturally-occurring gums, for example gum acacia or gum tragacanth,naturally-occurring phosphatides, for example soy bean, lecithin, andesters or partial esters derived from fatty acids and hexitolanhydrides, for example sorbitan monooleate, and condensation productsof the said partial esters with ethylene oxide, for examplepolyoxyethylene sorbitan monooleate. The emulsions may also containsweetening and flavoring agents.

Syrups and elixirs may be formulated with sweetening agents, for exampleglycerol, propylene glycol, sorbitol or sucrose. Such formulations mayalso contain a demulcent, a preservative and flavoring and coloringagents.

The pharmaceutical compositions may be in the form of a sterileinjectable aqueous or oleagenous suspension. This suspension may beformulated according to the known art using those suitable dispersing orwetting agents and suspending agents which have been mentioned above.The sterile injectable preparation may also be a sterile injectablesolution or suspension in a non-toxic parenterally-acceptable diluent orsolvent, for example as a solution in 1,3-butane diol. Among theacceptable vehicles and solvents that may be employed are water,Ringer's solution and isotonic sodium chloride solution. In addition,sterile, fixed oils are conventionally employed as a solvent orsuspending medium. For this purpose any bland fixed oil may be employedincluding synthetic mono- or diglycerides. In addition, fatty acids suchas oleic acid find use in the preparation of injectables.

The compounds of the present invention may also be administered in theform of suppositories for rectal administration of the drug. Thesecompositions can be prepared by mixing the drug with a suitablenon-irritating excipient which is solid at ordinary temperatures butliquid at the rectal temperature and will therefore melt in the rectumto release the drug. Such materials are cocoa butter and polyethyleneglycols.

For topical use, creams, ointments, jellies, solutions or suspensions,etc., containing the compounds of the present invention are employed. Asused herein, topical application is also meant to include the use ofmouth washes and gargles.

The pharmaceutical composition and method of the present invention mayfurther comprise other therapeutically active compounds as noted hereinwhich are usually applied in the treatment or prevention of the abovementioned pathological conditions.

Methods of Use

In yet another aspect, the present invention provides methods oftreating CXCR3-mediated conditions or diseases by administering to asubject having such a disease or condition, a therapeutically effectiveamount of a compound or composition of the invention. The “subject” isdefined herein to include animals such as mammals, including, but notlimited to, primates (e.g., humans), cows, sheep, goats, horses, dogs,cats, rabbits, rats, mice and the like.

As used herein, the phrase “CXCR3-mediated condition or disease” andrelated phrases and terms refer to a condition characterized byinappropriate, e.g., less than or greater than normal, CXCR3 activity.Inappropriate CXCR3 activity might arise as the result of CXCR3expression in cells which normally do not express CXCR3, increased CXCR3expression (leading to, e.g., inflammatory and immunoregulatorydisorders and diseases), or, decreased CXCR3 expression (leading to,e.g., certain cancers and angiogenic and vasculogenic-relateddisorders). Inappropriate CXCR3 functional activity might arise as theresult of CXCR3 expression in cells which normally do not express CXCR3,increased CXCR3 expression (leading to, e.g., inflammatory andimmunoregulatory disorders and diseases) or decreased CXCR3 expression.Inappropriate CXCR3 functional activity might also arise as the resultof chemokine secretion by cells which normally do not secrete a CXCchemokine, increased chemokine expression (leading to, e.g.,inflammatory and immunoregulatory disorders and diseases) or decreasedchemokine expression. A CXCR3-mediated condition or disease may becompletely or partially mediated by inappropriate CXCR3 functionalactivity. However, a CXCR3-mediated condition or disease is one in whichmodulation of CXCR3 results in some effect on the underlying conditionor disease (e.g., a CXCR3 antagonist results in some improvement inpatient well-being in at least some patients).

The term “therapeutically effective amount” means the amount of thesubject compound that will elicit the biological or medical response ofa tissue, system, animal or human that is being sought by theresearcher, veterinarian, medical doctor or other clinician or that issufficient to prevent development of or alleviate to some extent one ormore of the symptoms of the disease being treated.

Diseases and conditions associated with inflammation, infection andcancer can be treated with the present compounds and compositions. Inone group of embodiments, diseases or conditions, including chronicdiseases, of humans or other species can be treated with inhibitors ofCXCR3 function. These diseases or conditions include: (1) inflammatoryor allergic diseases such as systemic anaphylaxis or hypersensitivityresponses, drug allergies, insect sting allergies and food allergies;inflammatory bowel diseases, such as Crohn's disease, ulcerativecolitis, ileitis and enteritis; vaginitis; psoriasis and inflammatorydermatoses such as dermatitis, eczema, atopic dermatitis, allergiccontact dermatitis, urticaria; vasculitis; spondyloarthropathies;scleroderma; asthma and respiratory allergic diseases such as allergicrhinitis, hypersensitivity lung diseases, and the like, (2) autoimmunediseases, such as arthritis (rheumatoid and psoriatic), multiplesclerosis, systemic lupus erythematosus, type I diabetes,glomerulonephritis, and the like, (3) graft rejection (includingallograft rejection and graft-v-host disease) and conditions associatedtherewith, and (4) other diseases in which undesired inflammatoryresponses are to be inhibited, e.g., atherosclerosis, myositis,neurodegenerative diseases (e.g., Alzheimer's disease), encephalitis,meningitis, hepatitis, nephritis, sepsis, sarcoidosis, conjunctivitis,otitis, chronic obstructive pulmonary disease, sinusitis and Behcet'ssyndrome. In another group of embodiments, diseases or conditions aretreated with agonists of CXCR3 function. Examples of diseases to betreated with CXCR3 agonists include cancers, diseases in whichangiogenesis or neovascularization play a role (neoplastic diseases,retinopathy and macular degeneration), infectious diseases andimmunosuppressive diseases.

Preferably, the present methods are directed to the treatment orprevention of diseases or conditions selected from neurodegenerativediseases (e.g., Alzheimer's disease), multiple sclerosis, systemic lupuserythematosus, rheumatoid arthritis, atherosclerosis, encephalitis,meningitis, hepatitis, nephritis, sepsis, sarcoidosis, psoriasis,eczema, uticaria, type I diabetes, asthma, conjunctivitis, otitis,allergic rhinitis, chronic obstructive pulmonary disease, sinusitis,dermatitis, inflammatory bowel disease, ulcerative colitis, Crohn'sdisease, Behcet's syndrome, gout, cancer, viral infections (e.g., HIV),bacterial infections, and organ transplant conditions or skin transplantconditions. The term “organ transplant conditions” is meant to includebone marrow transplant conditions and solid organ (e.g., kidney, liver,lung, heart, pancreas or combination thereof) transplant conditions.

Diseases or conditions that can be treated with the present compoundsand compositions include diseases commonly associated with (I)inflammatory or allergic diseases, (2) autoimmune diseases, (3) graftrejection and (4) other diseases in which undesired inflammatoryresponses are to be inhibited, as described above. For example,restenosis following a procedure such as balloon angioplasty, iscommonly associated with atherosclerosis and can be treated with thepresent compounds and compositions.

Depending on the disease to be treated and the subject's condition, thecompounds of the present invention may be administered by oral,parenteral (e.g., intramuscular, intraperitoneal, intravenous, ICV,intracistemal injection or infusion, subcutaneous injection, orimplant), inhalation spray, nasal, vaginal, rectal, sublingual, ortopical routes of administration and may be formulated, alone ortogether, in suitable dosage unit formulations containing conventionalnon-toxic pharmaceutically acceptable carriers, adjuvants and vehiclesappropriate for each route of administration.

In the treatment or prevention of conditions which require chemokinereceptor modulation an appropriate dosage level will generally be about0.001 to 100 mg per kg patient body weight per day which can beadministered in single or multiple doses. Preferably, the dosage levelwill be about 0.01 to about 25 mg/kg per day; more preferably about 0.05to about 10 mg/kg per day. A suitable dosage level may be about 0.01 to25 mg/kg per day, about 0.05 to 10 mg/kg per day, or about 0.1 to 5mg/kg per day. Within this range the dosage may be 0.005 to 0.05, 0.05to 0.5 or 0.5 to 5.0 mg/kg per day. For oral administration, thecompositions are preferably provided in the form of tablets containing1.0 to 1000 milligrams of the active ingredient, particularly 1.0, 5.0,10.0, 15.0. 20.0, 25.0, 50.0, 75.0, 100.0, 150.0, 200.0, 250.0, 300.0,400.0, 500.0, 600.0, 750.0, 800.0, 900.0, and 1000.0 milligrams of theactive ingredient for the symptomatic adjustment of the dosage to thepatient to be treated. The compounds may be administered on a regimen of1 to 4 times per day, preferably once or twice per day.

It will be understood, however, that the specific dose level andfrequency of dosage for any particular patient may be varied and willdepend upon a variety of factors including the activity of the specificcompound employed, the metabolic stability and length of action of thatcompound, the age, body weight, general health, sex, diet, mode and timeof administration, rate of excretion, drug combination, the severity ofthe particular condition, and the host undergoing therapy.

The compounds of the present invention can be combined with othercompounds having related utilities to treat or prevent inflammatory andimmune disorders and diseases, including asthma and allergic diseases,as well as autoimmune pathologies such as rheumatoid arthritis andatherosclerosis, and those pathologies noted above. In many instances,compositions which include a compound of the invention and analternative or second therapeutic agent have additive or synergisticeffects when administered.

For example, in the treatment or prevention of inflammation, the presentcompounds may be used in conjunction or combination with anantiinflammatory or analgesic agent such as an opiate agonist, alipoxygenase inhibitor, such as an inhibitor of 5-lipoxygenase, acyclooxygenase inhibitor, such as a cyclooxygenase-2 inhibitor, aninterleukin inhibitor, such as an interleukin-1 inhibitor, an NMDAantagonist, an inhibitor of nitric oxide or an inhibitor of thesynthesis of nitric oxide, a non-steroidal antiinflammatory agent, or acytokine-suppressing antiinflammatory agent, for example with a compoundsuch as acetaminophen, aspirin, codiene, fentanyl, ibuprofen,indomethacin, ketorolac, morphine, naproxen, phenacetin, piroxicam, asteroidal analgesic, sufentanyl, sunlindac, tenidap, and the like.Similarly, the instant compounds may be administered with a painreliever; a potentiator such as caffeine, an H2-antagonist, simethicone,aluminum or magnesium hydroxide; a decongestant such as phenylephrine,phenylpropanolamine, pseudophedrine, oxymetazoline, ephinephrine,naphazoline, xylometazoline, propylhexedrine, or levo-desoxy-ephedrine;an antiutussive such as codeine, hydrocodone, caramiphen,carbetapentane, or dextromethorphan; a diuretic; and a sedating ornon-sedating antihistamine. Likewise, compounds of the present inventionmay be used in combination with other drugs that are used in thetreatment/prevention/suppression or amelioration of the diseases orconditions for which compounds of the present invention are useful. Suchother drugs may be administered, by a route and in an amount commonlyused therefor, contemporaneously or sequentially with a compound of thepresent invention. When a compound of the present invention is usedcontemporaneously with one or more other drugs, a pharmaceuticalcomposition containing such other drugs in addition to the compound ofthe present invention is preferred. Accordingly, the pharmaceuticalcompositions of the present invention include those that also containone or more other active ingredients, in addition to a compound of thepresent invention. Examples of other active ingredients that may becombined with a compound of the present invention, either administeredseparately or in the same pharmaceutical compositions, include, but arenot limited to: (a) VLA-4 antagonists, (b) steroids such asbeclomethasone, methylprednisolone, betamethasone, prednisone,dexamethasone, and hydrocortisone; (c) immunosuppressants such ascyclosporine (cyclosporine A, Sandimmune®, Neoral®), tacrolimus (FK-506,Prograf®), rapamycin (sirolimus, Rapamune®) and other FK-506 typeimmunosuppressants, and mycophenolate, e.g., mycophenolate mofetil(CellCept®); (d) antihistamines (H1-histamine antagonists) such asbromopheniramine, chlorpheniramine, dexchlorpheniramine, triprolidine,clemastine, diphenhydramine, diphenylpyraline, tripelennamine,hydroxyzine, methdilazine, promethazine, trimeprazine, azatadine,cyproheptadine, antazoline, pheniramine pyrilamine, astemizole,terfenadine, loratadine, cetirizine, fexofenadine,descarboethoxyloratadine, and the like; (e) non-steroidalanti-asthmatics such as .beta.2-agonists (terbutaline, metaproterenol,fenoterol, isoetharine, albuterol, bitolterol, and pirbuterol),theophylline, cromolyn sodium, atropine, ipratropium bromide,leukotriene antagonists (zafirlukast, montelukast, pranlukast,iralukast, pobilukast, SKB-106,203), leukotriene biosynthesis inhibitors(zileuton, BAY-1005); (f) non-steroidal antiinflammatory agents (NSAIDs)such as propionic acid derivatives (alminoprofen, benoxaprofen, bucloxicacid, carprofen, fenbufen, fenoprofen, fluprofen, flurbiprofen,ibuprofen, indoprofen, ketoprofen, miroprofen, naproxen, oxaprozin,pirprofen, pranoprofen, suprofen, tiaprofenic acid, and tioxaprofen),acetic acid derivatives (indomethacin, acemetacin, alclofenac, clidanac,diclofenac, fenclofenac, fenclozic acid, fentiazac, furofenac, ibufenac,isoxepac, oxpinac, sulindac, tiopinac, tolmetin, zidometacin, andzomepirac), fenamic acid derivatives (flufenamic acid, meclofenamicacid, mefenamic acid, niflumic acid and tolfenamic acid),biphenylcarboxylic acid derivatives (diflunisal and flufenisal), oxicams(isoxicam, piroxicam, sudoxicam and tenoxican), salicylates (acetylsalicylic acid, sulfasalazine) and the pyrazolones (apazone,bezpiperylon, feprazone, mofebutazone, oxyphenbutazone, phenylbutazone);(g) cyclooxygenase-2 (COX-2) inhibitors such as celecoxib (Celebrex®)and rofecoxib (Vioxx®); (h) inhibitors of phosphodiesterase type IV(PDE-IV); (i) gold compounds such as auranofin and aurothioglucose, (j)inhibitors of phosphodiesterase type IV (PDE-IV); (k) other antagonistsof the chemokine receptors, especially CCR1, CCR2, CCR3, CCR5, CCR6,CCR8 and CCR10; (1) cholesterol lowering agents such as HMG-CoAreductase inhibitors (lovastatin, simvastatin and pravastatin,fluvastatin, atorvastatin, and other statins), sequestrants(cholestyramine and colestipol), nicotinic acid, fenofibric acidderivatives (gemfibrozil, clofibrat, fenofibrate and benzafibrate), andprobucol; (m) anti-diabetic agents such as insulin, sulfonylureas,biguanides (metformin), α-glucosidase inhibitors (acarbose) andglitazones (troglitazone and pioglitazone); (n) preparations ofinterferon beta (interferon β-1 α, interferon β-1 β); (o) etanercept(Enbrel(®), (p) antibody therapies such as orthoclone (OKT3), daclizumab(Zenapax®), infliximab (Remicade®), basiliximab (Simulect®) andanti-CD40 ligand antibodies (eg., MRP-1); and (q) other compounds suchas 5-aminosalicylic acid and prodrugs thereof, hydroxychloroquine,D-penicillamine, antimetabolites such as azathioprene and6-mercaptopurine, and cytotoxic cancer chemotherapeutic agents. Theweight ratio of the compound of the present invention to the secondactive ingredient may be varied and will depend upon the effective doseof each ingredient. Generally, an effective dose of each will be used.Thus, for example, when a compound of the present invention is combinedwith an NSAID the weight ratio of the compound of the present inventionto the NSAID will generally range from about 1000:1 to about 1:1000,preferably about 200:1 to about 1:200. Combinations of a compound of thepresent invention and other active ingredients will generally also bewithin the aforementioned range, but in each case, an effective dose ofeach active ingredient should be used.

Immunosuppressants within the scope of the present invention furtherinclude, but are not limited to, leflunomide, RAD001, ERL080, FTY720,CTLA-4, antibody therapies such as orthoclone (OKT3), daclizumab(Zenapax®) and basiliximab (Simulect®), and antithymocyte globulins suchas thymoglobulins.

In particularly preferred embodiments, the present methods are directedto the treatment or prevention of multiple sclerosis using a compound ofthe invention either alone or in combination with a second therapeuticagent selected from betaseron, avonex, azathioprene (Imurek®, Imuran®),capoxone, prednisolone and cyclophosphamide. When used in combination,the practitioner can administer a combination of the therapeutic agents,or administration can be sequential.

In still other particularly preferred embodiments, the present methodsare directed to the treatment or prevention of rheumatoid arthritis,wherein the compound of the invention is administered either alone or incombination with a second therapeutic agent selected from the groupconsisting of methotrexate, sulfasalazine, hydroxychloroquine,cyclosporine A, D-penicillamine, infliximab (Remicade®), etanercept(Enbrel®), auranofin and aurothioglucose.

In yet other particularly preferred embodiments, the present methods aredirected to the treatment or prevention of an organ transplant conditionwherein the compound of the invention is used alone or in combinationwith a second therapeutic agent selected from the group consisting ofcyclosporine A, FK-506, rapamycin, mycophenolate, prednisolone,azathioprene, cyclophosphamide and an antilymphocyte globulin.

In yet another aspect, the present invention includes methods toevaluate putative specific agonists or antagonists of CXCR3 function.Accordingly, the present invention is directed to the use of thesecompounds in the preparation and execution of screening assays forcompounds which modulate the activity of the CXCR3 chemokine receptor.For example, the compounds of this invention are useful for isolatingreceptor mutants, which are excellent screening tools for more potentcompounds. Furthermore, the compounds of this invention are useful inestablishing or determining the binding site of other compounds to theCXCR3 chemokine receptor, e.g., by competitive inhibition. The compoundsof the instant invention are also useful for the evaluation of putativespecific modulators of the CXCR3 chemokine receptor, relative to otherchemokine receptors including CCR1, CCR2, CCR2A, CCR2B, CCR3, CCR4,CCR5, CCR6, CCR8, CCR10, CXCR3 and CXCR4. One of skill in the art willappreciate that thorough evaluation of specific agonists and antagonistsof the above chemokine receptors has been hampered by the lack ofavailability of non-peptidyl (metabolically resistant) compounds withhigh binding affinity for these receptors. Thus the compounds providedherein are particularly useful in this context. Combinatorial librariesof putative CXCR3 agonists or antagonists can be screened forpharmacological activity in in vitro or in vivo assays. Conventionally,new chemical entities with useful properties are generated byidentifying a chemical compound (called a “lead compound”) with somedesirable property or activity, e.g., CXCR3 chemokine receptormodulation activity, creating variants of the lead compound, andevaluating the property and activity of those variant compounds.However, the current trend is to shorten the time scale for all aspectsof drug discovery. Because of the ability to test large numbers quicklyand efficiently, high throughput screening (HTS) methods are replacingconventional lead compound identification methods.

In one preferred embodiment, high throughput screening methods involveproviding a library containing a large number of potential therapeuticcompounds (candidate compounds). Such “combinatorial chemical libraries”are then screened in one or more assays to identify those librarymembers (particular chemical species or subclasses) that display adesired characteristic activity. The compounds thus identified can serveconventional “lead compounds” or can themselves be used as potential oractual therapeutics.

A combinatorial chemical library is a collection of diverse chemicalcompounds generated by either chemical synthesis or biological synthesisby combining a number of chemical “building blocks” such as reagents.For example, a linear combinatorial chemical library, such as apolypeptide (e.g., mutein) library, is formed by combining a set ofchemical building blocks called amino acids in every possible way for agiven compound length (i.e., the number of amino acids in a polypeptidecompound). Millions of chemical compounds can be synthesized throughsuch combinatorial mixing of chemical building blocks (Gallop et. al.(1994) J. Med. Chem. 37(9):1233-1251).

Preparation and screening of combinatorial chemical libraries is wellknown to those of skill in the art. Such combinatorial chemicallibraries include, but are not limited to, peptide libraries (see, e.g.,U.S. Pat. No. 5,010,175, Furka (1991) Int. J. Pept. Prot. Res.37:487-493, Houghton et. al. (1991) Nature 354: 84-88), peptoidlibraries (PCT Publication No WO 91/19735), encoded peptide libraries(PCT Publication WO 93/20242), random bio-oligomer libraries (PCTPublication WO 92/00091), benzodiazepine libraries (U.S. Pat. No.5,288,514), libraries of diversomers, such as hydantoins,benzodiazepines and dipeptides (Hobbs et. al. (1993) Proc. Nat. Acad.Sci. USA 90:6909-6913), vinylogous polypeptide libraries (Hagihara etal. (1992) J. Amer. Chem. Soc. 114:6568), libraries of nonpeptidylpeptidomimetics with a Beta-D-Glucose scaffolding (Hirschmann et al.(1992) J. Amer. Chem. Soc. 114:9217-9218), analogous organic synthesesof small compound libraries (Chen et. al. (1994) J. Am. Chem. Soc.116:2661), oligocarbamate libraries (Cho et al. (1993) Science 261:1303)and/or peptidyl phosphonate libraries (Campbell et al. (1994) J. Org.Chem. 59:658). See, generally, Gordon et al. (1994) J. Med. Chem.37:1385-1401, nucleic acid libraries (see, e.g., Stratagene Corp.),peptide nucleic acid libraries (see, e.g., U.S. Pat. No. 5,539,083),antibody libraries (see, e.g., Vaughn et. al. (1996) NatureBiotechnology 14(3):309-314), and PCT/US96/10287), carbohydratelibraries (see, e.g., Liang et al. (1996) Science 274:1520-1522, andU.S. Pat. No. 5,593,853), and small organic molecule libraries (see,e.g., benzodiazepines, Baum (1993) C&EN Jan 18, page 33; isoprenoids,U.S. Pat. No. 5,549,974; pyrrolidines, U.S. Pat. Nos. 5,525,735 and5,519,134; morpholino compounds, U.S. Pat. No. 5,506,337;benzodiazepines, U.S. Pat. No. 5,288,514; and the like).

Devices for the preparation of combinatorial libraries are commerciallyavailable (see, e.g., 357 MPS, 390 MPS, Advanced Chem Tech, LouisvilleKy.; Symphony, Rainin, Woburn Mass.; 433A Applied Biosystems, FosterCity Calif.; 9050 Plus, Millipore, Bedford, Mass.).

A number of well known robotic systems have also been developed forsolution phase chemistries. These systems includes automatedworkstations like the automated synthesis apparatus developed by TakedaChemical Industries, LTD. (Osaka, Japan) and many robotic systemsutilizing robotic arms (Zymate II, Zymark Corporation, Hopkinton Mass.;Orca, Hewlett-Packard, Palo Alto Calif.), which mimic the manualsynthetic operations performed by a chemist. Any of the above devicesare suitable for use with the present invention. The nature andimplementation of modifications to these devices (if any) so that theycan operate as discussed herein will be apparent to persons skilled inthe relevant art. In addition, numerous combinatorial libraries arethemselves commercially available (see e.g., ComGenex, Princeton N.J.;Asinex, Moscow, Russia; Tripos, Inc., St. Louis Mo.; ChemStar, Ltd,Moscow, Russia; 3D Pharmaceuticals, Exton Pa.; Martek Biosciences,Columbia Md.; etc.).

High throughput assays for the presence, absence, quantification, orother properties of particular compounds may be used to test acombinatorial library that contains a large number of potentialtherapeutic compounds (potential modulator compounds). The assays aretypically designed to screen large chemical libraries by automating theassay steps and providing compounds from any convenient source toassays, which are typically run in parallel (e.g., in microtiter formatson microtiter plates in robotic assays). Preferred assays detectenhancement or inhibition of CXCR3 receptor function.

High throughput screening systems are commercially available (see e.g.,Zymark Corp., Hopkinton Mass.; Air Technical Industries, Mentor Ohio;Beckman Instruments, Inc., Fullerton Calif.; Precision Systems, Inc.,Natick Mass.; etc.). These systems typically automate entire procedures,including all sample and reagent pipetting, liquid dispensing, timedincubations, and final readings of the microplate in detector(s)appropriate for the assay. These configurable systems provide highthroughput and rapid start up as well as a high degree of flexibilityand customization. The manufacturers of such systems provide detailedprotocols for various high throughput systems. Thus, for example, ZymarkCorp. provides technical bulletins describing screening systems fordetecting the modulation of gene transcription, ligand binding, and thelike.

EXAMPLES

Reagents and solvents used below can be obtained from commercial sourcessuch as Aldrich Chemical Co. (Milwaukee, Wis., USA). ¹H-NMR spectra wererecorded on a Varian Gemini 400 MHz NMR spectrometer. Significant peaksare tabulated in the order: number of protons, multiplicity (s, singlet;d, doublet; t, triplet; q, quartet; m, multiplet; br s, broad singlet)and coupling constant(s) in Hertz (Hz). Electron Ionization (EI) massspectra were recorded on a Hewlett Packard 5989A mass spectrometer. Massspectrometry results are reported as the ratio of mass over charge,followed by the relative abundance of each ion (in parentheses). Intables, a single m/e value is reported for the M+H (or, as noted, M−H)ion containing the most common atomic isotopes. Isotope patternscorrespond to the expected formula in all cases. Electrospray ionization(ESI) mass spectrometry analysis was conducted on a Hewlett-Packard 1100MSD electrospray mass spectrometer using the HP1 100 HPLC for sampledelivery. Normally the analyte was dissolved in methanol at 0.1 mg/mLand 1 microliter was infused with the delivery solvent into the massspectrometer, which scanned from 100 to 1500 daltons. All compoundscould be analyzed in the positive ESI mode, using 1:1 acetonitrile/waterwith 1% acetic acid as the delivery solvent. The compounds providedbelow could also be analyzed in the negative ESI mode, using 2 mM NH₄OAcin acetonitrile/water as delivery solvent.

Example 1 Synthesis of Compound 1.01

The synthesis of compound 1.01 in six steps from commercially availableanthranilic acid provides an example of 3H-quinazolin4-one synthesis byMethod 1. Scheme 1 provides an overview of the synthetic route, forwhich the experimental details follow.

2-Propionylamino-benzoic acid (II). To a room temperature solution of50.22 g anthranilic acid (I) (370 mmol, 1.00 equiv) dissolved in 200 mLdry DMF was added 35.0 mL propionyl chloride (400 mmol, 1.10 equiv)dropwise by addition funnel over 1.5 h. The addition rate was slowenough to maintain internal temperature of the reaction below 38° C.Upon completed addition of the acid chloride, the heterogeneous reactionmixture was stirred for 2.5 h at ambient temperature and then pouredinto 1600 mL water. The resulting water/DMF mixture, with whiteprecipitate, was stirred vigorously at ambient temperature for one h,after which time the solid was collected by vacuum filtration, rinsingthe solid with cold water (2×100 mL). The product was dried in vacuoover phosphorous pentoxide overnight affording 48.04 g of a white solid.m.p. 120.1° C. ¹H NMR (CDCl₃) δ1.30 (t, 3H, J=7.4 Hz), 2.52 (q, 2H,J=7.4 Hz), 7.12 (t, 1H, J=7.2 Hz), 7.60 (t, 1H, J=7.1 Hz), 8.13 (d, 1H,J=6.3 Hz), 8.76 (d, 1H, J=7.8 Hz) ppm. MS (ESI⁻) 192.1 [M−H]⁻.

2-Ethyl-benzo[d][1,3]oxazin-4-one (III). A mixture of 46.66 g2-propionylamino-benzoic acid (II) (240 mmol, 1.00 equiv) suspended in180 mL acetic anhydride was heated to reflux (external temperature 170°to 180° C., oil bath) in a reaction vessel fitted with a distillationhead. Acetic acid was distilled from the reaction (b.p. 116 to 118° C.)over 1.5 to 2 h, after which time acetic anhydride began to distill(b.p. 136 to 138° C.). The reaction was equilibrated to room temperatureand acetic anhydride removed by vacuum distillation; a light yellowsolid resulted from concentration of the reaction solution. The solidwas triturated with hexane, collected by filtration (3×100 mL volumes ofhexane), and then dried in vacuo over phosphorous pentoxide to afford33.26 g of a light yellow solid. m.p. 83.9° C. ¹H NMR (CDCl₃) δ1.37 (t,3H, J=7.6 Hz), 2.73 (q, 2H, J=7.6 Hz), 7.49 (t, 1H, J₁=1.1 Hz, J₂=7.6Hz), 7.56 (d, 1H, J=8.4 Hz), 7.78 (t, 1H, J₁=1.5 Hz, J₂=7.2 Hz), 8.18(d, 1H, J=7.0 Hz) ppm. MS (ESI⁺) 176.1 [MH]⁺.

2-Ethyl-3-(4-fluorophenyl)-3H-quinazolin-4-one (IV) A solution of 8.502-ethyl-benzo[d]1,3]oxazin-4-one (III) (48.5 mmol, 1.00 equiv) and 6.27g 4-fluoroaniline (50.9 mmol, 1.05 equiv) dissolved in 35 mL chloroformwas heated to reflux for 12 h, after which time TLC indicated nocompound III remained (R_(f)=0.51, 20% acetone in hexane). Thechloroform was removed in vacuo and the resulting solid suspended in 18mL ethylene glycol. A catalytic amount of sodium hydroxide (86 mg, 2.2mmol, 0.045 equiv) was added to the mixture, which was heated to 140 to150° C. (external temperature, oil bath). After 10 h, the reaction wasremoved from heat and equilibrated to room temperature; upon cooling aprecipitate formed. The cooled reaction product mixture was acidifiedwith 2 mL aqueous 5% hydrochloric acid solution and suspended in 20 mLcold water. The solid was collected by vacuum filtration, rinsing withcold water (2×50 mL) and cold isopropyl alcohol (2×50 mL). The air-driedsolid was recrystallized from isopropyl alcohol, affording 10.62 gtan-white needles. m.p. 178.3° C. ¹H NMR (CDCl₃) δ1.25 (t, 3H, J=7.4Hz), 2.46 (q, 2H, J=7.4 Hz), 7.26 (d, 2H, J=6.4 Hz), 7.27 (d, 2H, J=6.4Hz), 7.48 (t, 1H, J=6.8 Hz), 7.73-7.81 (m, 2H), 8.27 (d, 1H, J=7.96 Hz)ppm. MS (ESI⁺) 269.1 [MH]⁺.

2-(1-Bromoethyl)-3-(4-fluorophenyl)-3H-quinazolin-4-one (V). To asolution of 7.084 g 2-ethyl-3-(4-fluorophenyl)-3H-quinazoline-4-one (IV)(26.40 mmol, 1.000 equiv) and 2.60 g sodium acetate (31.7 mmol, 1.20equiv) dissolved in 30 mL glacial acetic acid at 40° C. (externaltemperature; oil bath) was added dropwise by addition funnel a solutionof 1.36 mL bromine (26.4 mmol, 1.00 equiv) in 5 mL glacial acetic acidover 60 min. Upon completed addition of the bromine solution, thereaction was stirred an additional 60 min, after which time TLCindicated no IV remained (R_(f)=0.44; 40% ethyl acetate in hexane) andthe heterogeneous mixture was poured into 400 mL water. The resultingaqueous, acidic mixture, with precipitate, was stirred vigorously atambient temperature for two h. The precipitate was collected by vacuumfiltration, rinsing with warm (ca. 40° C.) water (2×50 mL) and coldisopropyl alcohol (50 mL). The solid was dried in vacuo over phosphorouspentoxide overnight, affording 8.81 g of a white solid.m.p. 179.8° C. ¹HNMR (CDCl₃) δ2.06 (d, J=0.016p, 3H), 4.55 (q, 0.016p, 2H), 7.16 (ddd,1H, J₁=2.4 Hz, J₂=4.8 Hz, J₃=8.4 Hz), 7.24 (dt, 1H, J₁=2.8 Hz, J₂=8.0Hz), 7.28 (dt, ₁H, J₁=2.8 Hz, J₂=8.4 Hz), 7.51-7.58 (m, 2H), 7.80-7.81(m, 2H), 8.28 (dt, 1H, J₁=0.8 Hz, J₂=8.0 Hz) ppm. MS (ESI⁺) 348.0 [MH]⁺.

3-(4-Fluorophenyl)-2-[1-(2-methoxy-ethylamino)-ethyl]-3H-quinazolin-4-one(VI). A solution of 242 mg from2-(1-bromoethyl)-3-(4-fluorophenyl)-3H-quinazolin-4-one (V) (0.697 mmol,1.00 equiv) and 160 μL 1-amino-2-methoxyethane (1.81 mmol, 2.60 equiv)in 5 mL absolute ethanol was heated to reflux for 26 h then concentratedin vacuo to remove the ethanol. The resulting yellow foam waspartitioned between dichloromethane and aqueous saturated sodiumbicarbonate solution (25 mL each). The separated aqueous layer wasextracted again with dichloromethane (20 mL). Combined organic extractswere dried over sodium sulfate, filtered, and concentrated in vacuo toyield a yellow foam. The crude product was purified by chromatography onsilica gel (3.5 cm o.d.×12 cm h) eluting with 5% methanol in chloroform.Fractions containing product at R_(f)=0.31, 5% methanol in chloroform,were combined and concentrated in vacuo to afford 220 mg product as alight yellow foam. ¹H NMR (CDCl₃) δ1.26 (d, 3H, J=6.4 Hz), 2.35 (br s,1H), 2.54 (ddd, 1H, J₁=4.4 Hz, J₂=6.0 Hz, J₃=10.4 Hz), 2.71 (ddd, 1H,J₁=4.0 Hz, J₂=7.2 Hz, J₃=11.2 Hz), 3.27 (s, 3H), 3.36-3.45 (m, 2H), 3.47(q, 1H, J=6.4 Hz), 7.22-7.26 (m, 4H), 7.46 (ddd, 1H, J₁=1.6 Hz, J₂=6.8Hz, J₃=8.0 Hz), 7.71-7.78 (m, 2H), 8.25 (dd, 1H, J₁=1.2 Hz, J₂=8.0 Hz)ppm. MS (ESI⁺) 342.2 [MH]⁺

Compound 1.01. To a solution of 130 mg3-(4-fluorophenyl)-2-[1-(2-methoxy-ethylamino)-ethyl]-3H-quinazolin-4-one(VI) (0.381 mmol, 1.00 equiv), 59 μL triethylamine (0.419 mmol, 1.10equiv), and 2 mg DMAP (16 μmol, 0.04 equiv) dissolved in 3 mL1,4-dioxane at room temperature was added 79 μL neat decanoyl chloride(0.381 mmol, 1.00 equiv); a colorless precipitate developed. Thereaction mixture was stirred overnight at room temperature thenconcentrated in vacuo to remove the dioxane. The resulting concentratewas partitioned between dichloromethane and aqueous saturated sodiumbicarbonate solution (20 mL each). The separated aqueous layer wasextracted again with dichloromethane (15 mL) and the combined organicextracts dried over sodium sulfate, filtered, and concentrated in vacuoto yield a yellow, glassy oil. The crude product was purified bychromatography on silica gel (2.5 cm o.d.×10 cm h) eluting with agradient of 20 to 25% ethyl acetate in hexane. Fractions containingproduct at R_(f)=0.84, 5% methanol in chloroform, were combined andconcentrated in vacuo to afford 120 mg of a colorless solid. m.p. 71.4°C. ¹H NMR (d₆-DMSO; T=140° C.) δ0.90 (t, 3H, J=7.2 Hz), 1.18-1.44 (m,14H), 1.44 (d, 3H, J=7.2 Hz), 1.98-2.08 (m, 2H), 3.11 (s, 3H), 3.33-3.52(m, 4H), 5.11 (br q, 1H, J=6.0 Hz), 7.32 (br m, 3H), 7.49 (br m, 1H),7.55 (ddd, 1H, J₁=1.2 Hz, J₂=7.6 Hz, J₃=8.0 Hz), 7.73 (d, 1H, J=8.0 Hz),7.85 (ddd, 1H, J₁=1.2 Hz, J₂=7.2 Hz, J₃=8.4 Hz), 8.15 (dd, 1H, J₁=1.6Hz, J₂=8.0 Hz) ppm. At room temperature, compound exists as a mixture ofcis/trans amide rotamers, ca. 1:1 determined by integration ofcharacteristic ¹H NMR peaks (CDCl₃, T=25° C.) at δ_(minor) 4.78 (q,1.0H, J=7.2 Hz) and δ_(major) 5.33 (q, 1.2H, J=7.2 Hz) ppm. MS (ESI⁺)496.4 [MH]⁺

Synthesis of Compound 1.02

Compound 1.02 was prepared following the synthesis of compound 1.01described above. Method 1 was followed for the synthetic sequence,wherein 1-(2-aminoethyl)pyrrolidine was used in step e instead of2-(dimethylamino)-1-aminoethane. Characterization data for compound 1.02follows: colorless, viscous oil. ¹H NMR similar to spectrum for compound1.01: a mixture of cis/trans amide rotamers in ca. 2:1 (CDCl₃; T=25° C.)characteristic resonance peaks at δ_(minor) 4.78 (q, 1.0H, J=6.8 Hz) andδ_(major) 5.33 (q, 1.8H, J=7.6 Hz) ppm. MS (ESI⁺) 535.4 [MH]⁺

Synthesis of Compound 1.03

Compound 1.03 was prepared following the synthesis of compound 1.01described above. Method 1 was followed for the synthetic sequence,wherein 1-(2-Aminoethyl)morpholine was used in step e instead of2-(dimethylamino)-1-aminoethane. Characterization data for compound 1.03follows: colorless, viscous oil. ¹H NMR (d₆-DMSO; T=140° C.) δ0.89 (t,3H, J=6.8 Hz), 1.18-1.46 (m, 14H), 1.46 (d, 3H, J=6.4 Hz), 1.98-2.08 (m,2H), 2.23-2.34 (m, 5H), 2.47 (ddd, 1H, , J₁=6.0 Hz, J₂=8.8 Hz, J₃=14.4Hz), 3.31 (ddd, 1H, , J₁=5.6 Hz, J₂=8.4 Hz, J₃=14.4 Hz), 3.39-3.49 (m,5H), 5.10 (br q, 1H), 7.32 (br m, 3H), 7.51 (br m, 1H), 7.56 (ddd, 1H,J₁=0.8 Hz, J₂=J₃=8.0 Hz), 7.72 (d, 1H, J=7.6 Hz), 7.86 (ddd, 1H, J₁=1.6Hz, J₂=7.2 Hz, J₃=8.4 Hz), 8.15 (dd, 1H, J₁=0.8 Hz, J₂=7.2 Hz) ppm. Atroom temperature, compound exists as a mixture of cis/trans amide, ca.4:3 (CDCl₃; T=25° C.) characteristic resonance peaks at δ_(minor) 4.77(q, 1.0H, J=6.4 Hz) and δ_(major) 5.33 (q, 1.3H, J=6.8 Hz) ppm. MS(ESI⁺) 551.5 [MH]⁺

Synthesis of Compound 1.04

Compound 1.04 was prepared following the synthesis of compound 1.01described above. Method 1 was followed for the synthetic sequence,wherein 5-(2-Aminoethyl)imidazole was used in step e instead of2-(dimethylamino)-1-aminoethane. Characterization data for compound 1.04follows: colorless, viscous oil. ¹H NMR similar to spectrum for compound1.01: a mixture of cis/trans amide rotamers in ca. 3:1 (CDCl₃; T=25° C.)characteristic resonance peaks at δ_(minor) 4.81 (q, 1.0H, J=6.8 Hz) andδ_(major) 5.05 (q, 2.7H, J=7.2 Hz) ppm. MS (ESI⁺) 532.3 [MH]⁺.

Synthesis of Compound 1.05

Compound 1.05 was prepared following the synthesis of compound 1.01described above. Method 1 was followed for the synthetic sequence,wherein biphenylacetyl chloride was used in step f instead of decanoylchloride. Characterization data for compound 1.05 follows: yellow,viscous oil. ¹H NMR similar to spectrum for compound 1.01: a mixture ofcis/trans amide rotamers in ca. 2:1 (CDCl₃; T=25° C.) characteristicresonance peaks at δ_(minor) 4.89 (q, 1.0H, J=6.8 Hz) and δ_(major) 5.32(q, 1.8H, J=6.8 Hz) ppm. MS (ESI⁺) 549.2 [MH]⁺

Synthesis of Compound 1.06

Compound 1.06 was prepared following the synthesis of 1.01 describedabove. Method 1 was followed for the synthetic sequence, whereinbiphenylcarbonyl chloride was used in step f instead of decanoylchloride. Characterization data for compound 1.06 follows: white solid.m.p. =147.3° C. ¹H NMR similar to spectrum for compound 1.01: a mixtureof cis/trans amide rotamers in ca. 3:1 (CDCl₃; T=25° C.) determined byintegration of characteristic resonance peaks at δ_(minor) 5.02 (br q,1.0H) and δ_(major) 5.43 (br q, 3.0H) ppm. MS (ESI⁺) 535.2 [MH]⁺

Synthesis of Compound 1.07

Compound 1.07 was prepared following the synthesis of 1.01 describedabove. Method 1 was followed for the synthetic sequence, wherein3-(3-Aminopropyl)-(3H)-imidazole was used in step e instead of2-(Dimethylamino)-1-aminoethane. Characterization data for compound 1.07follows: colorless, viscous oil. ¹H NMR similar to spectrum for compound1.01: a mixture of cis/trans amide rotamers in ca. 1:1 (CDCl₃; T=25° C.)determined by integration of characteristic resonance peaks at δ_(minor)4.77 (q, 1.0H, J=6.8 Hz) and δ_(major) 5.28 (q, 1.1H, J=7.6 Hz) ppm. MS(ESI⁺) 546.3 [MH]⁺

Synthesis of Compound 1.08

Compound 1.08 was prepared following the synthesis of compound 1.01described above. Method 1 was followed for the synthetic sequence,wherein 1-(3-Aminopropyl)morpholine was used in step e instead of2-(Dimethylamino)-1-aminoethane. Characterization data for compound 1.08follows: pale yellow glass. ¹H NMR similar to spectrum for compound1.01: a mixture of cis/trans amide rotamers in ca. 2:1 (CDCl₃; T=25° C.)determined by integration of characteristic resonance peaks at minor4.77 (q, 1.0H, J=6.4 Hz) and δ_(minor) 5.38 (q, 1.8H, J=7.2 Hz) ppm. MS(ESI⁺) 565.4 [MH]⁺

Synthesis of Compound 1.09

Compound 1.09 was prepared following the synthesis of 1.01 describedabove. Method 1 was followed for the synthetic sequence, whereinbiphenylcarbonyl chloride was used in step f instead of decanoylchloride. Characterization data for compound 1.09 follows: white solid.m.p.=153.0° C. ¹H NMR (d₆-DMSO; T=140° C.) δ1.42 (d, 3H, J=7.2 Hz), 2.07(s, 6H), 2.26 (ddd, 1H, J₁=5.6 Hz, J₂=9.2 Hz, J₃12.4 Hz), 2.46 (ddd, 1H,J₁=5.2 Hz, J₂=9.2 Hz, J₃=14.4 Hz), 3.36 (d, 1H, J=15.2 Hz), 3.38 (ddd,1H, J₁=5.2 Hz, J₂=8.8 Hz, J₃=14.8 Hz), 3.49 (ddd, 1H, J₁=6.0 Hz, J₂9.2Hz, J₃=15.2 Hz), 3.50 (d, 1H, J=15.2 Hz), 5.15 (q, 1H, J=6.8 Hz), 7.12(d, 2H, J=7.6 Hz), 7.20 (t, 1H, J=7.2 Hz), 7.26 (dd, 2H, J₁=7.2 Hz,J₂=7.6 Hz), 7.36 (br m, 3H), 7.53 (br m, 1H), 7.56 (ddd, 1H, J₁=1.2 Hz,J₂=7.2 Hz, J₃=8.0 Hz), 7.72 (d, 1H, J=7.2 Hz), 7.87 (ddd, 1H, J₁=1.6 Hz,J₂=7.2 Hz, J₃=8.4 Hz), 8.16 (dd, 1H, J₁=1.6 Hz, J₂ 8.0 Hz) ppm. At roomtemperature, compound exists as a mixture of cis/trans amide rotamers,ca. 2:1 (CDCl₃; T=25° C.) determined by integration of characteristicresonance peaks at δ_(minor) 4.84 (q, 1.0H, J=6.8 Hz) and δ_(major) 5.30(q, 2.1H, J=6.8 Hz) ppm. MS (ESI⁺) 473.3 [MH]⁺

Synthesis of Compound 1.10

Compound 1.10 was prepared following the synthesis of 1.01 describedabove. Method 1 was followed for the synthetic sequence, wherein5-(2-Aminoethyl)imidazole was used in step e instead of2-(Dimethylamino)-1-aminoethane. Characterization data for compound 1.10follows: yellow, viscous oil. ¹H NMR similar to spectrum for compound1.01: a mixture of cis/trans amide rotamers in ca. 3:2 (CDCl₃; T=25° C.)determined by integration of characteristic resonance peaks at δ_(minor)4.77 (q, 1.0H, J=6.8 Hz) and δ_(major) 5.37 (q, 1.6H, J=6.8 Hz) ppm. MS(ESI⁺) 577.4 [MH]⁺

Synthesis of Compound 1.11

Compound 1.11 was prepared following the synthesis of compound 1.01described above. Method 1 was followed for the synthetic sequence,wherein (4-methylphenyl)acetyl chloride was used in step f instead ofdecanoyl chloride. Characterization data for compound 1.11 follows:white solid. m.p. 188.3° C. ¹H NMR similar to spectrum for compound1.09: a mixture of cis/trans amide rotamers in ca. 2:1 (CDCl₃; T=25° C.)determined by integration of characteristic resonance peaks at δ_(minor)5.02 (q, 1.0H, J=6.8 Hz) and δ_(major) 5.47 (q, 1.9H, J=7.2 Hz) ppm. MS(ESI⁺) 487.3 [MH]⁺

Synthesis of Compound 1.12

Compound 1.12 was prepared following the synthesis of compound 1.01described above. Method 1 was followed for the synthetic sequence,wherein (4-bromophenyl)acetyl chloride was used in step f instead ofdecanoyl chloride.

Characterization data for compound 1.12 follows: colorless glass. ¹H NMRsimilar to spectrum for compound 1.09: a mixture of cis/trans amiderotamers in ca. 2:1 CDCl₃; T=25° C.) determined by integration ofcharacteristic resonance peaks at δ_(minor) 4.82 (q, 1.0H, J=7.2 Hz) andδ_(major) 5.27 (q, 2.3H, J=6.8 Hz) ppm. MS (ESI⁺) 551.2 [MH]⁺

Synthesis of Compound 1.13

To a solution of 112 mg zinc(II) bromide (500 μmol, 10 equiv) at 0° C.was added 1.0 mL 1-propenylmagnesium bromide solution in 0.5 mnL THF(0.5 M; 500 μmol, 10 equiv). The resulting white, cloudy mixture wasstirred at 0° C. for 60 min before a solution of 27 mg 1.12 (49 μmol,1.0 equiv) and 4 mg bis-dppf palladium(II) dichloride (5 μmol, 0.1equiv) dissolved in 0.5 mL THF was added all at once by cannulation. Thereaction mixture was stirred at room temperature for 14 h, then heatedto 60° C. (external temperature, oil bath) to drive the reaction towardcompletion. After 2 h at 60° C., 5 mL saturated aqueous ammoniumchloride solution was added to the cooled (0° C.) reaction mixture. Theaqueous layer was extracted with ethyl acetate (3×15 mL) and thecombined organic separations dried over magnesium sulfate, filtered, andconcentrated in vacuo to yield a yellow film. The crude product waspurified by flash column chromatography on silica gel (3.5 cm o.d.×10 cmh) eluting with 5% methanol in chloroform to yield 8 mg product olefinas a colorless film. The product was isolated as a mixture of olefinisomers, which were separated by preparative HPLC (reverse phase,CH₃CN:H₂O). Compound 1.13 eluted before the trans olefin isomer 1.14. ¹HNMR similar to spectrum for compound 1.09: a mixture of cis/trans amiderotamers in ca. 2:1 (CDCl₃; T=25° C.) determined by integration ofcharacteristic resonance peaks at δ_(major) 4.85 (q, 1.9H, J=6.8 Hz) andδ_(minor) 5.13 (q, 1.0H, J=7.2 Hz) ppm. MS (ESI⁺) 513.2 [MH]⁺

Synthesis of Compound 1.14

Compound 1.14 was prepared coincidentally with compound 1.12 andisolated by preparative HPLC as the second product to elute. ¹H NMRsimilar to spectrum for compound 1.09: a mixture of cis/trans amiderotamers in ca. 2:1 (CDCl₃; T=25° C.) determined by integration ofcharacteristic resonance peaks at δ_(major) 4.83 (q, 1.8H, J=7.2 Hz) andδ_(minor) 5.12 (q, 1.0H, J=7.6 Hz) ppm. MS (ESI⁺) 513.2 [MH]⁺

Synthesis of Compound 1.15

Hydrogen gas was introduced by balloon to a nitrogen-purged, evacuatedflask charged with 4.8 mg 1.13 and 1.14 (9.4 μmol, 1.0 equiv) and 5.0 mgpalladium on activated carbon (10% wt Pd; 4.7 μmol, 0.5 equiv) suspendedin 2.0 mL methanol at room temperature. The reaction was stirred at roomtemperature for 18 h then filtered through a pad of celite. The filtratewas concentrated in vacuo then purified by column chromatography onsilica gel (2.0 cm o.d.×8 cm h) eluting with 5% methanol in chloroform.Fractions containing product were concentrated in vacuo to afford 4.5 mgof a colorless film. ¹H NMR similar to spectrum for compound 1.09: amixture of cis/trans amide rotamers in ca. 3:2 (CDCl₃; T=25° C.)determined by integration of characteristic resonance peaks at δ_(major)4.83 (q, 1.4H, J=6.8 Hz) and δ_(minor) 5.20 (q, 1.0H, J=7.2 Hz) ppm. MS(ESI⁺) 515.3 [MH]⁺

Synthesis of Compound 1.16

A degassed (3×freeze-evacuate-thaw cycles) biphasic mixture of 27.0 mg1.12 (49.0 μmol, 1.00 equiv), 34.0 mg 4-fluorophenylboronic acid (245μmol, 5.00 equiv), and 3.0 mg tetrakistriphenylphosphine palladium(0)(2.5 μmol, 0.05 equiv) in 3.0 mL toluene and 3.0 mL 2M aqueous sodiumcarbonate was heated to 100° C. (external temperature, oil bath). After4 h, MS indicated no compound 1.12 remained and the separated aqueouslayer was extracted with 50% ethylacetate in hexane (2×15 mL). Combinedorganic extracts were dried over magnesium sulfate, filtered, andconcentrated in vacuo to yield a yellow oil. The crude material waspurified by chromatography on silica gel (3.5 cm o.d.×12 cm h) elutingwith 5% methanol in chloroform. Fractions containing product werecombined and concentrated in vacuo to afford 27.0 mg product as acolorless, viscous oil. ¹H NMR similar to spectrum for compound 1.09: amixture of cis/trans amide rotamers in ca. 3:2 (CDCl₃; T=25° C.)determined by integration of characteristic resonance peaks at δ_(major)4.90 (q, 1.3H, J=7.2 Hz) and δ_(minor) 5.30 (q, 1.0H, J=7.2 Hz) ppm. MS(ESI⁺) 567.2 [MH]⁺.

Synthesis of Compound 1.17

Compound 1.17 was prepared following the synthesis of 1.01 describedabove. Method 1 was followed for the synthetic sequence, wherein2-methoxy-1-aminoethane was used in step e instead of2-(N,N-dimethylamino)-1-aminoethane, and biphenylacetyl chloride wasused in step f instead of decanoyl chloride. Characterization data forcompound 1.17 follows: beige solid. m.p.=153.8° C. ¹H NMR similar tospectrum for compound 1.09: a mixture of cis/trans amide rotamers in ca.2:1 (CDCl₃; T=25° C.) determined by integration of characteristicresonance peaks at δ_(major) 4.89 (q, 1.0H, J=6.4 Hz) and δ_(minor) 5.33(q, 1.8H, J=6.8 Hz) ppm. MS (ESI⁺) 536.2 [MH]⁺

Synthesis of Compound 1.18

Compound 1.18 was prepared following the synthesis of compound 1.01described above. Method 1 was followed for the synthetic sequence,wherein 1-(2-aminoethyl)morpholine was used in step e instead of2-(N,N-dimethylamino)-1-aminoethane, and biphenylacetyl chloride wasused in step f instead of decanoyl chloride. Characterization data forcompound 1.18 follows: colorless, viscous oil. ¹H NMR similar tospectrum for compound 1.09: a mixture of cis/trans amide rotamers in ca.2:1 (CDCl₃; T=25° C.) determined by integration of characteristicresonance peaks at δ_(minor) 4.88 (q, 1.0H, J=6.8 Hz) and δ_(major) 5.32(q, 1.7H, J=7.2 Hz) ppm. MS (ESI⁺) 591.3 [MH]⁺

Synthesis of Compound 1.19

Compound 1.19 was prepared following the synthesis of compound 1.01described above. Method 1 was followed for the synthetic sequence,wherein 2-ethoxy-1-aminoethane was used in step e instead of2-(N,N-dimethylamino)-1-aminoethane, and biphenylacetyl chloride wasused in step f instead of decanoyl chloride. Characterization data forcompound 1.19 follows: light yellow, glassy solid. m.p.=150.6° C. ¹H NMR(d₆-DMSO; T=140° C.) δ0.98 (t, 3H, J=6.8 Hz), 1.43 (d, 3H, J=6.8 Hz),3.29-3.63 (m, 8H), 5.18 (q, 1H, J=6.0 Hz), 7.20 (d, 2H, J=7.6 Hz),7.27-7.36 (m, 3H), 7.41-7.47 (m, 3H), 7.49-7.64 (m, 6H), 7.72 (d, 1H,J=8.0 Hz), 7.85 (ddd, 1H, J₁=1.6 Hz, J₂=8.2 Hz, J₃=8.6 Hz), 8.15 (d, 1H,J=8.0 Hz) ppm. At room temperature, compound exists as a mixture ofcis/trans amide rotamers, ca. 2:1 (CDCl₃; T=25° C.) determined byintegration of characteristic resonance peaks at δ_(minor) 4.87 (q,1.0H, J=6.8 Hz) and δ_(major) 5.33 (q, 2.1H, J=7.2 Hz) ppm. MS (ESI⁺)550.2 [MH]⁺

Synthesis of Compound 1.20

Compound 1.20 was prepared following the synthesis of compound 1.01described above. Method 1 was followed for the synthetic sequence,wherein 3-aminopropionitrile was used in step e instead of2-(N,N-dimethylanino)-1-aminoethane, and biphenylacetyl chloride wasused in step f instead of decanoyl chloride. Characterization data forcompound 1.20 follows: colorless glass. ¹H NMR similar to spectrum forcompound 1.19: a mixture of cis/trans amide rotamers in ca. 1:1 (CDCl₃;T=25° C.) determined by integration of characteristic resonance peaks atδ_(A) 4.94 (q, 1.0H, J=6.8 Hz) and δ_(B) 5.14 (q, 1.0H, J=7.6 Hz) ppm.MS (ESI⁺) 530.2 [MH]⁺

Synthesis of Compound 1.21

Compound 1.21 was prepared following the synthesis of compound 1.01described above. Method 1 was followed for the synthetic sequence,wherein 2-isopropoxy -1-aminoethane was used in step e instead of2-(N,N-dimethylamino)-1-aminoethane, and biphenylacetyl chloride wasused in step f instead of decanoyl chloride. Characterization data forcompound 1.21 follows: faint yellow glass. ¹H NMR similar to spectrumfor compound 1.19: a mixture of cis/trans amide rotamers in ca. 3:1(CDCl₃; T=25° C.) determined by integration of characteristic resonancepeaks at δ_(minor) 4.88 (q, 1.0H, J=6.7 Hz) and δ_(major) 5.30 (q, 2.9H,J=7.0 Hz) ppm. MS (ESI⁺) 564.2 [MH]⁺

Synthesis of Compound 1.22

Compound 1.22 was prepared following the synthesis of compound 1.01described above. Method 1 was followed for the synthetic sequence,wherein 2-aminomethyl pyridine was used in step e instead of2-(N,N-dimethylamino)-1-aminoethane, and biphenylacetyl chloride wasused in step f instead of decanoyl chloride. Characterization data forcompound 1.22 follows: colorless glass. ¹H NMR similar to spectrum forcompound 1.19: a mixture of cis/trans amide rotamers in ca. 1:1 (CDCl₃;T=25° C.) determined by integration of characteristic resonance peaks atδ_(A) 5.13 (q, 1.0H, J=6.4 Hz) and δ_(B) 5.46 (q, 1.0H, J=8.0 Hz) ppm.MS (ESI⁺) 569.3 [MH]⁺

Synthesis of Compound 1.23

Compound 1.23 was prepared following the synthesis of compound 1.01described above. Method 1 was followed for the synthetic sequence,wherein 2-aminomethyl pyridine was used in step e instead of2-(N,N-dimethylamino)-1-aminoethane, and biphenylacetyl chloride wasused in step f instead of decanoyl chloride. Characterization data forcompound 1.23 follows: colorless glass. ¹H NMR similar to spectrum forcompound 1.19: a mixture of cis/trans amide rotamers in ca. 1:1 (CDCl₃;T=25° C.) determined by integration of characteristic resonance peaks atδ_(A) 5.13 (q, 1.0H, J=6.4 Hz) and δ_(B) 5.46 (q, 1.0H, J=8.0 Hz) ppm.MS (ESI⁺) 569.3 [MH]⁺

Synthesis of Compound 1.24

Compound 1.24 was prepared following the synthesis of compound 1.01described above. Method 1 was followed for the synthetic sequence,wherein 3-(3-aminopropyl)imidazole was used in step e instead of2-(N,N-dimethylamino)-1-aminoethane, and biphenylacetyl chloride wasused in step f instead of decanoyl chloride. Characterization data forcompound 1.24 follows: colorless oil. ¹H NMR similar to spectrum forcompound 1.19: a mixture of cis/trans amide rotamers in ca. 1:1 (CDCl₃;T=25° C.) determined by integration of characteristic resonance peaks atδ_(A) 4.89 (q, 1.0H, J=6.6 Hz) and δ_(B) 5.29 (q, 1.1H, J=7.1 Hz) ppm.MS (ESI⁺) 569.3 [MH]⁺.

Synthesis of Compound 1.25

To a mixture of 175 mg 1.19 (318 μmol, 1.00 equiv) and 500 mg zincpowder (7.65 mmol, 24.0 equiv) suspended in 3.0 mL glacial acetic acidat 40° C. (external temperature, oil bath) was added ca. 200 μLconcentrated aqueous hydrochloric acid (5 drops by pipet, 18 M; 3.6mmol). The resulting beige, cloudy reaction mixture evolved gas and wasstirred at 40° C. for 15 min, then decanted from the suspendedsolids/zinc and neutralized with concentrated aqueous sodium hydroxideto pH>12. The aqueous, alkaline solution was extracted withdichloromethane (3×20 mL). Combined organic extracts were dried overmagnesium sulfate, filtered, and concentrated in vacuo to yield acolorless oil. The crude material was purified by chromatography onsilica gel (3.5 cm o.d.×10 cm h) eluting with 2% methanol in chloroform.Fractions containing product at R_(f)=0.52, 10% methanol in chloroform,were combined and concentrated in vacuo to afford 83 mg of a colorlessoil. ¹H NMR similar to spectrum for compound 1.19: a mixture ofcis/trans amide rotamers in ca. 2:1 (CDCl₃; T=25° C.) determined byintegration of characteristic resonance peaks at δ_(minor) 4.62 (q,1.0H, J=7.1 Hz) and δ_(major) 5.31 (q, 2.1H, J=7.0 Hz) ppm. MS (ESI⁺)536.3 [MH]⁺

Synthesis of Compound 1.26

Compound 1.26 was prepared following the synthesis of compound 1.01described above. Method 1 was followed for the synthetic sequence,wherein 2-ethoxy-1-aminoethane was used in step e instead of2-(N,N-dimethylamnino)-1-aminoethane, and(4-trifluoromethylphenyl)acetic acid was used, with EDC and catalyticHOBT, in step f instead of decanoyl chloride. Characterization data forcompound 1.26 follows: colorless oil. ¹H NMR similar to spectrum forcompound 1.19: a mixture of cis/trans amide rotamers in ca. 5:2 (CDCl₃;T=25° C.) determined by integration of characteristic resonance peaks atδ_(minor) 4.85 (q, 1.0H, J=6.8 Hz) and δ_(major) 5.33 (q, 2.6H, J=6.8Hz) ppm. MS (ESI⁺) 542.2 [MH]⁺

Synthesis of Compound 1.27

Compound 1.27 was prepared following the synthesis of compound 1.01described above. Method 1 was followed for the synthetic sequence,wherein 3-methylaminopyridine was used in step e instead of2-(N,N-dimethylarnino)-1-aminoethane, and(4-trifluoromethylphenyl)acetic acid was used, with EDC and catalyticHOBT, in step f instead of decanoyl chloride. Characterization data forcompound 1.27 follows: colorless oil. ¹H NMR similar to spectrum forcompound 1.19: a mixture of cis/trans amide rotamers in ca. 6:5 (CDCl₃;T=25° C.) determined by integration of characteristic resonance peaks atδ_(minor) 4.99 (q, 1.0H, J=6.6 Hz) and δ_(major) 5.37 (q, 1.2H, J=7.2Hz) ppm. MS (ESI⁺) 561.2 [MH]⁺

Synthesis of Compound 1.28

Compound 1.28 was prepared following the synthesis of compound 1.01.MS(ESI⁺) 533.3, 534.3. ¹H NMR (DMSO, T=140° C.) 0.87 (t, 3H, J=7.0 Hz),1.26 (m, 14H), 1.66 (m, 4H), 2.22 (m, 2H), 2.49-2.76 (m, 6H), 3.51 (t,2H, J=3.3 Hz), 3.87 (s, 3H), 4.24 (s, 2H), 7.11 (m, 2H), 7.31 (m, 2H),7.51 (m, 1H), 7.60 (m, 1H), 7.80 (m, 1H), 8.13 (m, 1H). MS(ESI⁺) 533.8(M⁺).

Synthesis of Compound 1.29

Compound 1.29 was prepared following the synthesis of compound 1.01.Colorless viscous oil; mixture of cis /trans amide rotamers (1/1),determined by ¹H NMR (CDCl₃) 4.82 (q, 1H, J=7.5 Hz), 5.37 (q, 1H, J=7.5Hz). MS(ESI⁺) 547.2 (MH⁺). Anal. (C₂₃H₂₈N₄O₂) cal. C, 72.49; H, 8.48; N10.25. Found C, 72.62; H, 8.44; N, 10.12.

Synthesis of Compound 1.30

Compound 1.30 was prepared following the synthesis of 1.01. Yellowsolid. Mixture of cis/trans amide rotamers(1/1), determined by ¹H NMR(CDCl₃) 1.40 (d, 3H, J=6.8 Hz), 1.46 (d, 3H, J=6.8 Hz). MS(ESI⁺) 561.2(MH⁺).

Synthesis of Compound 1.31

Compound 1.31 was prepared following the synthesis of 1.01. Colorlessviscous oil; mixture of cis/trans amide rotamers (1/1), determined by ¹HNMR (CDCl₃) 4.88(q, 1H, J=7.2 Hz), 5.38 (q, 1H, J=7.2 Hz). MS(ESI⁺)575.5 (MH⁺). Anal. (C₃₅H₅₀N₄O₃) cal. C, 73.14; H, 8.77; N, 9.75. FoundC, 72.45; H, 8.75; N, 9.08.

Synthesis of Compound 1.32

Compound 1.32 was prepared following the synthesis of 1.01. Colorlessviscous oil; mixture of cis/trans amide rotamers (2/3), determined by ¹HNMR (CDCl₃) 4.87(q, 1H, J=7.2 Hz), 5.38 (q, 1H, J=7.2 Hz). MS(ESI⁺)522.3 (MH⁺). Anal. (C₃₁H₄₃N₃O₄) cal. C, 71.37; H, 8.31; N, 8.05. FoundC, 71.13; H, 8.42; N, 8.02.

Synthesis of Compound 1.33

Compound 1.33 was prepared following the synthesis of 1.01. Yellowsolid. m.p. 96.9° C. mixture of cis/trans amide rotamers (1/1),determined by ¹H NMR (CDCl₃) 4.87(q, 1H, J=7.2 Hz), 5.38 (q, 1H, J=7.2Hz). MS(ESI⁺) 605.3 (MH⁺). Anal. (C₃₇H₃₇FN₄O₃C₄H₈O₂) cal. C, 71.08; H,6.55; N 8.09. Found C, 71.96; H, 6.19; N, 8.47.

Synthesis of Compound 1.34

Compound 1.34 was prepared following the synthesis of 1.01. white solid.m.p. 116.3° C. mixture of cis/trans amide rotamers (1/1), determined by¹H NMR (CDCl₃) 4.96(q, 1H, J=7.2 Hz), 5.38 (q, 1H, J=7.2 Hz). MS(ESI⁺)587.3 (MH⁺). Anal. (C₃₇H₃₈N₄O₃) cal. C, 75.74; H, 6.53; N, 9.55. FoundC, 75.05; H, 6.56; N, 9.35.

Synthesis of Compound 1.35

Compound 1.35 was prepared following the synthesis of 1.01. yellowsolid. Mixture of cis/trans amide rotamers (3/8), determined by ¹H NMR(CDCl₃) 4.89(m, 1H), 5.38 (m, 1H). MS(ESI⁺) 575.3 (MH⁺). Anal.(C₃₆H₃₅FN₄O₂ C₄H₈O₂) cal. C, 72.49; H, 6.54; N, 8.45. Found C, 72.77; H,6.10; N, 8.89.

Synthesis of Compound 1.36

Compound 1.36 was prepared following the synthesis of 1.01. white solid;m.p. 61.3° C. mixture of cis/trans amide rotamers (1/1), determined by¹H NMR (CDCl₃) 4.92(q, 1H, J=7.2 Hz), 5.32 (q, 1H, J=7.2 Hz).MS(ESI⁺)591.3 (MH⁺). Anal. (C₃₆H₃₈N₄O₄) cal. C, 73.20; H, 6.48; N, 9.48. FoundC, 72.92; H, 6.46; N, 9.29.

Synthesis of Compound 1.37

Compound 1.37 was prepared following the synthesis of 1.01. white solid;mixture of cis/trans amide rotamers (1/2), determined by ¹H NMR (CDCl₃)4.86(q, 1H, J=7.2 Hz), 5.32 (q, 1H, J=7.2 Hz).MS(ESI⁺) 515.3 (MH⁺).Anal. (C₃₁H₃₅FN₄O₂) cal C, 72.35; H, 6.85; N, 10.89. Found C, 72.11; H,6.92; N, 10.71.

Synthesis of Compound 1.38

Compound 1.38 was prepared following the synthesis of 1.01. white solid;mixture of cis/trans amide rotamers (1/1.7), determined by ¹H NMR(CDCl₃) 4.85 (q, 1H, J=7.2 Hz), 5.30 (q, 1H, J=7.2 Hz).Ms(ESI⁺) 545.3(MH⁺). Anal. (C₃₂H₃₇FN₄O₃) cal. C, 70.57; H, 6.85; N, 10.29. Found C,70.33; H, 6.90; N, 10.13.

Synthesis of Compound 1.39

Compound 1.39 was prepared following the synthesis of 1.01. white solid;mixture of cis/trans amide rotamers (1/1), determined by ¹H NMR (CDCl₃)4.95 (q, 1H, J=7.2 Hz), 5.32 (q, 1H, J=7.2 Hz).MS(ESI⁺) 562.3 (MH⁺).Anal. (C₃₅H₃₅N₃O₄) cal. C, 74.84; H, 6.28; N, 7.48. Found C, 74.56; H,6.26; N, 7.30.

Synthesis of Compound 1.40

Compound 1.40 was prepared following the synthesis of 1.01. White solid;mixture of cis/trans amide rotamers (2/1), determined by ¹H NMR (CDCl₃)4.70 (m, 1H), 5.38 (t, 1H, J=7.0 Hz).MS(ESI⁺) 604.3 (MH⁺). Anal.(C₃₈H₄₁N₃O₄) cal. C, 75.60; H, 6.84; N, 6.96. Found C, 74.98; H, 6.82;N, 6.72.

Synthesis of Compound 1.42

Compound 1.42 was prepared following the synthesis of 1.01. white solid;mixture of cis/trans amide rotamers (1/2), determined by ¹H NMR (CDCl₃)4.86 (q, 1H, J=7.3 Hz), 5.30 (q, 1H, J=7.3 Hz).MS(ESI⁺) 644.2 (MH⁺).Anal. (C₃₃H₃₀IN₃O₃) cal. C, 61.59; H, 4.70; N, 6.53. Found C, 61.63; H,4.73; N, 6.36.

Synthesis of Compound 1.43

The mixture of 1.42 (1 mmol, 0.643 g) and CuCN (3 mmol, 0.27 g) in 0.10ml of DMF was heated to 130° C. for 10 h. After evaporating the solvent,the residue was dissolved in CH₂Cl₂, the organic layer was washed bywater, brine, dried over NaSO₄ and removed in vacuo to give a sticky oilwhich was purified by chromatography to afford a white solid; mixture ofcis/trans amide rotamers (1/2), determined by ¹H NMR (CDCl₃) 4.75 (q,1H, J=7.3 Hz), 5.28 (q, 1H, J=7.3 Hz).MS(ESI⁺) 543.2 (MH⁺). Anal.(C₃₄H₃₀N₄O₃) cal. C, 75.26; H, 5.57; N, 10.32. Found C, 75.00; H, 5.59;N, 10.19.

Synthesis of Compound 1.44

Compound 1.44 was prepared following the synthesis of 1.01. White solid.¹H NMR (CDCl₃) 1.40 (d, 3H, J=7.3 Hz), 3.05 (m, 1H), 3.12 (s, 3H), 3.25(m, 1H), 3.55-3.70 (m, 2H), 3.77 (d, 1H, J=15 Hz), 3.9 (d, 1H, J=15 Hz),5.08 (d, 1H, 12 Hz), 5.88 (m, 2H), 7.28-7.35 (m, 5H), 7.42 (m, 2H), 7.57(m, 5H), 7.72 (m, 1H), 7.80(m, 1H), 8.32(m, 1H), 8.55 (m, 2H). MS(ESI⁺)533.3 (MH⁺).

Synthesis of Compound 1.45

The mixture of 1.43 (0.1 mmol, 0.054 g) and 30% H₂O₂ (0.6 mmol) in 1 mLof DMF and 1 ml of dioxane was stirred at room temperature for 1 h.Usual work up gave the give a white solid; mixture of cis/trans amiderotamers (1/2), determined by ¹H NMR(CDCl₃) 4.95 (q, 1H, J=7.3 Hz), 5.15(q, 1H, J=7.3 Hz).MS(ESI⁺) 561.3(MH⁺). Anal. (C₃₄H₃₂N₄O₄.C₄H₈O) cal. C,70.35; H, 6.21; N, 8.64. Found C, 70.98; H, 5.99; N, 9.14.

Synthesis of Compound 1.47

Compound 1.47 was prepared following the synthesis of 1.01, mixture ofcis/trans amide rotamers (1/17), determined by ¹H NMR (CDCl₃) 1.35 (d,3H, J=7.3 Hz), 1.42(d, 3H, J=7.3 Hz).MS(ESI⁺) 534.2 (MH⁺). Anal.(C₃₃H₃₁N₃O₄) cal. C, 74.28; H, 5.86; N, 7.87. Found C, 73.83; H, 5.93;N, 7.73.

Synthesis of Compound 1.48

Compound 1.48 was prepared following the synthesis of 21. white solid,mixture of cis/trans arnide rotamers (1/5), determined by ¹H NMR (CDCl₃)4.72 (q, 1H, J=7.0 Hz), 5.25 (q, 1H, J=7.0 Hz).MS(ESI⁺) 557.3 (MH⁺).Anal. (C₃₅H₃₂N₄O₃) cal. C, 75.52; H, 5.79; N, 10.06. Found C, 75.03; H,5.92; N, 9.96

Synthesis of Compound 1.49

Compound 1.49 was prepared following the synthesis of 1.01, white solid,m.p. 98.1° C., mixture of cis/trans amide rotamers (1/1), determined by¹H NMR (CDCl₃) 4.72 (q, 1H, J=7.0 Hz), 5.25 (q, 1H, J=7.0 Hz).MS(ESI⁺)576.3 (MH⁺). Anal. (C₃₆H₃₇N₃O₄) cal. C, 75.11; H, 6.48; N, 7.30. FoundC, 75.08; H, 6.59; N, 7.27.

Synthesis of Compound 1.50

Compound 1.50 was prepared following the synthesis of 1.01, white solid,¹H NMR (CDCl₃) 1.24 (d, 3H, J=6.8 Hz), 1.46 (t, 3H, J=6.9 Hz), 3.64 (s,2H), 4.09 (q, 2H, J=6.9 Hz), 4.83 (m, 1H), 6.90 (m, 1H), 7.05 (m, 2H),7.17 (m, 1H), 7.35-7.61 (m, 7H), 7.63 (m, 5H), 8.25 (m, 1H).MS(ESI⁺)504.2 (MH⁺). Anal. (C₃₂H₂₉N₃O₃) cal. C, 76.32; H, 5.80; N, 8.34. FoundC, 75.85; H, 5.88; N, 8.14

Synthesis of Compound 1.51

Compound 1.51 was prepared following the synthesis of 1.01, white solid,¹H NMR (CDCl₃) 1.05 (t, 3H, J=7.0 Hz), 1.40 (t, 3H, J=6.92 Hz), 2.81 (m,2H), 3.18 (m, 2H ), 3.80 (m, 2H), 3.91 (d, 1H, J=15 Hz), 4.0 (m, 2H),4.03 (d, 1H, J=15 Hz), 6.11 (m, 1H), 6.42 (m, 1H), 6.47 (m, 1H), 7.01(m, 3H), 7.22-7.58 (m, 14H), 7.64 (m, 1H), 7.75 (m, 1H), 8.27 (d, 1H,J=8 Hz). MS(ESI⁺) 638.3 (MH⁺). Anal. (C₄₁H₃₉N₃O₄) cal. C, 77.21; H,6.16; N, 6.59. Found C, 77.28; H, 6.15; N, 6.58.

Synthesis of Compound 1.52

Compound 1.53 was synthesized in a manner similar to that used for thesynthesis of 1.01. Under N₂, the mixture of pyridine-4-boronic acid(0.053 g, 0.43 mmol), 1.52 (0.050 g, 0.087 mmol) and Pd(PPh₃)₄ (0.010 g,0.009 mmol) in toluene (4 mL) and 3M Na₂CO₃ (4 mL) was heated to 110° C.for 3 h. The organic layer was washed with water, dried over NaSO₄ andevaporated to give a oil which was purified by chromatography to affordcompound 1.53 as a white solid (15 mg). MS(ESI⁺) 577.3 (MH⁺).

Synthesis of Compound 1.54

Under N₂, the mixture of 3,4-difluorophenylboronic acid (0.131 g, 0.83mmol), compound 1.52 (0.050 g, 0.087 mmol) and Pd(OAc)₂(0.016 g, 0.071mmol) in DME (3 mL) and 3M Na₂CO₃ (2 mL) was heated to 90° C. for 3 h.The aqueous layer was extracted with CH₂Cl₂, the combined organicextracts was dried over Na₂SO₄, filtered and concentrated. The residuewas purified by chromatography to give a white solid (71 mg,). ¹H NMR(DMSO, T=140° C.) 0.96 (d, 3H, J=6.8 Hz), 1.36 (t, 3H, J=7.2 Hz), 1.42(d, 3H, J=6.4 Hz), 3.31-3.56 (m, 8H), 4.13 (q, 2H, J=6.8 Hz), 5.16 (q,1H, J=6.4 Hz), 7.05 (br, 2H), 7.18-7.61 (mn 10H), 7.70 (d, 1H, J=8 Hz),7.84 (t, 1H, J=6.8 Hz), 8.13 (d, 1H, J=8.4 Hz). At room temperature,mixture of cis/trans amide rotamers (1/1), determined by ¹H NMR (CDCl₃)4.95 (q, 1H, J=6.8 Hz), 5.35 (q, 1H, J=6.8 Hz). MS(ESI⁺) 612.2(MH⁺).Anal. (C₃₆H₃₅F₂N₃O₄) cal. C, 70.69; H, 5.77; N, 6.87. Found C, 70.22; H,5.71; N, 6.81.

Synthesis of Compound 1.55

Trifluoroacetic anhydride (0.024 g, 0.113 mmol) was added dropwise to amixture of the amine (0.036 g, 0.094 mmol) and Et₃N (0.014 g, 0.142mmol) in CH₂Cl₂ at room temperature. After stirring for 1 h, the organiclayer was washed by water, brine, dried over NaSO₄ and removed in vacuoto give a oil which was purified by chromatography to afford a colorlessoil, compound 1.55. ¹H NMR (CDCl₃) 1.06 (t, 3H, J=7.04 Hz), 1.45(t, 3H,J=7.0 Hz), 1.54(d, 3H, J=7 Hz), 3.38 (m, 2H), 3.58 (t, 2H, J=6.2 Hz),3.70 (t, 2H, J=6.2 Hz), 4.08 (m, 2H), 5.19 (q, 1H, J=7 Hz), 7.01 (m,2H), 7.14 (m, 1H), 7.32 (m, 1H), 7.51(m, 1H), 7.77(m, 2H), 8.27 (d, 1H,J=7.3 Hz) MS(ESI⁺) 478.3 (MH⁺).

Example 2 Synthesis of Compound 2.01

The synthesis of 2.01 in five steps from commercially available 2-amino-6-methyl-benzoic acid is an example of 3H-quinazolin-4-one synthesis byMethod 2 (see Scheme 2, below).

2-Methyl-6-propionylamino-benzoic acid (VIII). To a room temperaturesolution of 4.35 g 2-amino-6-methyl-benzoic acid (VII) (28.8 mmol, 1.00equiv) dissolved in 25 mL dry DMF was added 2.75 mL propionyl chloride(31.7 mmol, 1.10 equiv) dropwise by addition funnel over 30 min. Uponcompleted addition of the acid chloride, the heterogeneous reactionmixture was stirred for 3 h at room temperature and then poured into 200mL water. The resulting water/DMF mixture, with white precipitate, wasstirred vigorously at ambient temperature for one h, after which timethe solid was collected by vacuum filtration, rinsing the solid withcold water (2×50 mL). The white solid was dried in vacuo overphosphorous pentoxide overnight to afford 4.65 g of a white solid. m.p.152.5° C. ¹H NMR (d₆-DMSO) δ1.06 (t, 3H, J=7.6 Hz), 2.29 (q, 2H, J=7.6Hz), 2.35 (s, 3H), 7.04 (d, 1H, J=7.6 Hz), 7.30 (dd, 1H, J₁=7.6 Hz,J₂=8.0 Hz), 7.47 (d, 1H, J=8.0 Hz), 9.57 (s, 1H), 13.18 (br s, 1H) ppm.MS (ESI⁻) 206.1 [M−H]⁻.

2-Ethyl-3-(4-fluoro-phenyl)-5-methyl-3H-quinazolin-4-one (IX)). To amixture of 4.266 g 2-methyl-6-propionylamino-benzoic acid (VIII) (20.58mmol, 1.00 equiv) and 2.14 mL 4-fluoroaniline (22.6 mmol, 1.10 equiv)suspended in 35 mL toluene was added a solution of 1.08 mL phosphoroustrichloride (12.3 mmol, 0.598 equiv) dissolved in 10 mL toluene dropwiseby addition funnel over 30 min. The resulting heterogeneous mixture washeated to reflux for 20 h and then cooled to room temperature anddiluted with 100 mL toluene. To the room temperature reaction mixturewas added 100 mL aqueous 10% sodium carbonate solution and the resultingbiphase was stirred vigorously until all solids dissolved. The toluenewas removed in vacuo and a precipitate developed. The solid wascollected by filtration, rinsing with water (2×75 mL). The air-driedsolid was purified by recrystallization from isopropyl alcohol to afford3.31 g colorless flakes, dried in vacuo over phosphorous pentoxide. m.p.170.0° C. ¹H NMR (CDCl₃) δ1.24 (t, 3H, J=7.6 Hz), 2.44 (q, 2H, J=7.6Hz), 2.84 (s, 3H), 7.25 (dd, J₁ =1.6 Hz, J₂=6.4 Hz), 7.27(2×d, 2×2H,J=6.4 Hz), 7.58 (dd, 1H, J₁=1.2 Hz, J₂=8.0 Hz), 7.63 (dd, 1H, J₁=J₂=8.0Hz) ppm. MS (ESI⁺) 283.2 [MH]⁺.

2-(1-Bromo-ethyl)-3-(4-fluoro-phenyl)-5-methyl-3H-quinazolin-4-one (X).To a mixture of 1.969 g2-ethyl-3-(4-fluoro-phenyl)-5-methyl-3H-quinazolin-4-one (IX) (6.974mmol, 1.000 equiv) and 0.687 g sodium acetate (8.37 mmol, 1.20 equiv) in28 mL glacial acetic acid at 40° C. (external temperature, oil bath) wasadded a solution of 0.372 mL bromine (7.32 mmol, 1.05 equiv) dissolvedin 5 mL glacial acetic acid dropwise by addition funnel over 30 min.After 2 h the reaction solution was poured into 250 mL water. Theresulting mixture was stirred vigorously at room temperature for 1 h,after which time the precipitate was collected by vacuum filtration,rinsing with warm (ca. 40° C.) water (3×50 mL). The solid was dried invacuo over phosphorous pentoxide overnight, affording 2.19 g of a whitesolid. m.p. decomposes upon heating. ¹H NMR (CDCl₃) δ2.04 (d, 3H, J=6.8Hz), 2.82 (s, 3H), 4.51 (q, 1H, J=6.8 Hz), 7.15 (ddd, 1H, J₁=2.4 Hz,J₂=4.4 Hz, J₃=8.4 Hz), 7.23 (dd, 1H, J₁=2.8 Hz, J₂=10.8 Hz), 7.25-7.31(m, 2H), 7.56 (ddd, J₁=2.8 Hz, J₂=4.8 Hz, J₃=8.8 Hz), 7.64 (2×d, 2×1H,J=5.2 Hz) ppm. MS (ESI⁺) 361.1 [MH]⁺.

3-(4-Fluoro-phenyl)-5-methyl-2-[1-(2-morpholin-4-yl-ethylamino)-ethyl]-3H-quinazolin-4-one(XI). A mixture of 0.283 g 2-(1-bromo-ethyl)-3-(4-fluoro-phenyl)-5-methyl-3H-quinazolin-4-one (X) (0.784 mmol, 1.00 equiv) and 0.165 mL1-(2-aminoethyl)morpholine (1.25 mmol, 1.60 equiv) in 5 mL ethanol washeated to reflux. After 20 h, the ethanol was removed in vacuo and theconcentrate partitioned between dichloromethane and saturated aqueoussodium bicarbonate solution (20 mL ea.). The separated aqueous layer wasextracted again with dichloromethane (15 mL) and the combined organicextracts dried over magnesium sulfate, filtered, and concentrated invacuo to yield a yellow foam. The crude material was purified bychromatography on silica gel (3.5 cm o.d.×12 cm h) eluting with 5%methanol in chloroform. Fractions containing product were combined andconcentrated in vacuo to afford 257 mg of a pale yellow solid. m.p.192.9° C. ¹H NMR (CDCl₃) δ1.27 (d, 3H, J=6.4 Hz), 2.26-2.34 (m, 3H),2.38-2.44 (m, 1H), 2.46-2.52 (m, 2H), 2.56-2.70 (m, 2H), 2.82 (s, 3H),3.39 (q, 1H, J=6.4 Hz), 3.70-3.80 (m, 4H), 7.18-7.29 (m, 5H), 7.46 (dd,1H, J₁=0.8 Hz, J₂=8.0 Hz), 7.61 (dd, 1H, J₁=7.6 Hz, J₂=7.8 Hz), ppm. MS(ESI⁺) 411.2 [MH]⁺

Compound 2.01. To a room temperature solution of 127 mg3-(4-fluoro-phenyl)-5-methyl-2-[1-(2-morpholin-4-yl-ethylamino)-ethyl]-3H-quinazolin-4-one(XI) (0.309 mmol, 1.00 equiv), 0.084 mL triethylamine (0.618 mmol, 2.00equiv), and 2.0 mg DMAP (0.016 mmol, 0.052 equiv) dissolved in 3 mLdichloromethane was added 107 mg biphenylacetyl chloride (0.463 mmol,1.50 equiv). The clear, faint yellow-colored reaction mixture wasstirred for 12 h at room temperature then poured into 10 mL saturatedaqueous sodium bicarbonate solution. The separated aqueous layer wasextracted with a second volume of dichloromethane (20 mL). The combinedorganic extracts dried over magnesium sulfate, filtered, andconcentrated in vacuo to yield an orange oil. The crude product waspurified by chromatography on silica gel (3.5 cm o.d.×10 cm h) elutingwith 2% methanol in chloroform. Fractions containing product atR_(f)=0.48, 5% methanol in chloroform, were combined and concentrated invacuo to afford 115 mg product as a faint yellow, viscous oil. ¹H NMR(d₆-DMSO; T=140° C.) 67 1.44 (d, 3H, J=6.4 Hz), 2.28-2.42 (m, 5H),2.50-2.60 (m, 1H), 2.77 (s, 3H), 3.38-3.64 (m, 8H), 5.12 (q, 1H, J=6.8Hz), 7.20 (m, 2H), 7.27-7.38 (m, 5H), 7.40-7.47 (m, 3H), 7.51-7.56 (m,3H), 7.58-7.64 (m, 2H), 7.68 (dd, 1H, J₁=J₂=7.6 Hz) ppm. At roomtemperature, compound exists as a mixture of cis/trans amide rotamers,ca. 3:2 by ¹H NMR (CDCl₃; T=25° C.) δ4.84 (q, 1.0H, J=6.8 Hz) & 5.28 (q,1.4H, J=6.8 Hz) ppm. MS (ESI⁺) 605.3 [MH]⁺

Synthesis of Compound 2.02

Compound 2.02 was prepared following the synthesis of 2.01 describedabove. Method 2 was followed for the synthetic sequence, wherein2-amino-3-methoxy-benzoic acid was used in step a instead of2-amino-6-methylbenzoic acid. Characterization data for compound 2.02follows: colorless, viscous oil. ¹H NMR similar to spectrum for compound2.01 a mixture of cis/trans amide rotamers in ca. 3:1 (CDCl₃; T=25° C.)characteristic resonance peaks at δ_(minor) 4.89 (q, 1.0H, J=6.8 Hz) andδ_(major) 5.28 (q, 2.8H, J=7.6 Hz) ppm. MS (ESI⁺) 579.3 [MH]⁺

Synthesis of Compound 2.03

Compound 2.03 was prepared following the synthesis of compound 2.01described above. Method 2 was followed for the synthetic sequence,wherein 2-amino-3-chloro-benzoic acid was used in step a instead of2-amino-6-methylbenzoic acid. Characterization data for compound 2.03follows: colorless, viscous oil. 1H NMR similar to spectrum for compound2.01: a mixture of cis/trans amide rotamers in ca. 3:1 (CDCl₃; T=25° C.)characteristic resonance peaks at δ_(minor) 4.89 (q, 1.0H, J=6.4 Hz) andδ_(major) 5.23 (q, 2.7H, J=6.8 Hz) ppm. MS (ESI⁺) 625.3 [MH]⁺

Synthesis of Compound 2.04

Compound 2.04 was prepared following the synthesis of compound 2.01described above. Method 2 was followed for the synthetic sequence,wherein 2-amino-3-methyl-benzoic acid was used in step a instead of2-amino-6-methyl-benzoic acid. Characterization data for compound 2.04follows: colorless, viscous oil. ¹H NMR similar to spectrum for compound2.01: a mixture of cis/trans amide rotamers in ca. 3:2 (CDCl₃; T=25° C.)characteristic resonance peaks at δ_(minor) 4.92 (q, 1.0H, J=6.7 Hz) andδ_(major) 5.35 (q, 1.7H, J=7.3 Hz) ppm. MS (ESI⁺) 605.3 [MH]⁺

Synthesis of Compound 2.05

Compound 2.05 was prepared following the synthesis of compound 2.01described above. Method 2 was followed for the synthetic sequence,wherein 2-amino-6-chloro-benzoic acid was used in step a instead of2-amino-6-methyl-benzoic acid. Characterization data for compound 2.05follows: colorless, viscous oil. ¹H NMR similar to spectrum for compound2.01: a mixture of cis/trans amide rotamers in ca. 2:1 (CDCl₃; T=25° C.)characteristic resonance peaks at δ_(minor) 4.84 (q, 1.0H, J=6.8 Hz) andδ_(major) 5.21 (q, 2.0H, J=6.8 Hz) ppm. MS (ESI⁺) 625.3 [MH]⁺

Synthesis of Compound 2.06

Compound 2.06 was prepared following the synthesis of compound 2.01described above. Method 2 was followed for the synthetic sequence,wherein 2-amino-6-fluoro-benzoic acid was used in step a instead of2-amino-6-methyl-benzoic acid and 2-ethoxy -1-aminoethane was used instep d instead of 1-(2-aminoethyl)morpholine. Characterization data forcompound 2.06 follows: colorless, viscous oil. ¹H NMR similar tospectrum for compound 2.01: a mixture of cis/trans amide rotamers in ca.5:2 (CDCl₃; T=25° C.) characteristic resonance peaks at δ_(minor) 4.87(q, 1.0H, J=6.7 Hz) and δ_(major) 5.27 (q, 2.5H, J=7.0 Hz) ppm. MS(ESI⁺) 568.2 [MH]⁺

Synthesis of Compound 2.07

Compound 2.07 was prepared following the synthesis of compound 2.01described above. Method 2 was followed for the synthetic sequence,wherein 2-ethoxy-1-aminoethane was used in step d instead of1-(2-aminoethyl)morpholine. Characterization data for compound 2.07follows: colorless, viscous oil. ¹H NMR similar to spectrum for compound2.01: a mixture of cis/trans amide rotamers in ca. 2:1 (CDCl₃; T=25° C.)characteristic resonance peaks at δ_(minor) 4.85 (q, 1.0H, J=6.8 Hz) andδ_(major) 5.29 (q, 1.8H, J=6.6 Hz) ppm. MS (ESI⁺) 564.3 [MH]⁺

Synthesis of Compound 2.08

Compound 2.08 was prepared following the synthesis of compound 2.01described above. Method 2 was followed for the synthetic sequence,wherein 4-ethoxyaniline was used in step b instead of 4-fluoroanilineand 2-ethoxy-1-aminoethane was used in step d instead of1-(2-aminoethyl)morpholine. Characterization data for compound 2.08follows: colorless, viscous oil. ¹H NMR similar to spectrum for compound2.01: a mixture of cis/trans amide rotamers in ca. 1:1 (CDCl₃; T=25° C.)characteristic resonance peaks at δ_(A) 4.95 (q, 1.1H, J=6.8 Hz) and 8B5.35 (q, 1.0H, J=6.8 Hz) ppm. MS (ESI⁺) 590.3 [MH]⁺

Synthesis of Compound 2.09

Compound 2.09 was prepared following the synthesis of compound 2.01described above. Method 2 was followed for the synthetic sequencewherein butyryl chloride was used in step a instead of propionylchloride, 4-cyanoaniline was used in step b instead of 4-fluoroaniline,and 2-ethoxy-1-aminoethane was used in step d instead of1-(2-aminoethyl)morpholine. Characterization data for compound 2.09follows: colorless, viscous oil. ¹H NMR similar to spectrum for compound2.01: a mixture of cis/trans amide rotamers in ca. 4:1 (CDCl₃; T=25° C.)characteristic resonance peaks at δ_(minor) 4.39 (dd, 1.0H, J₁=4.4 Hz,J₂=10.0 Hz) and δ_(major) 5.31 (dd, 3.9H, J₁=J₂=7.2 Hz ppm. MS (ESI⁺)585.3 [MH]⁺.

Synthesis of Compound 2.10

Compound 2.10 was prepared following the synthesis of compound 2.01described above. Method 2 was followed for the synthetic sequence,wherein 2-aminonicotinic acid was used in step a instead of2-amino-6-methylbenzoic acid, 4-ethoxyaniline was used in step b insteadof 4-fluoroaniline, and 2-ethoxy-1-aminoethane was used in step dinstead of 1-(2-aminoethyl)morpholine. Characterization data forcompound 2.10 follows: light yellow, viscous oil. ¹H NMR similar tospectrum for compound 2.01: a mixture of cis/trans amide rotamers in ca.1:1 (CDCl₃; T=25° C.) characteristic resonance peaks at δ_(minor) 5.04(q, 1.0H, J=6.4 Hz) and δ_(major) 5.41 (q, 1.0H, J=7.2 Hz) ppm. MS(ESI⁺) 577.3 [MH]⁺

Synthesis of Compound 2.11

Compound 2.11 was prepared following the synthesis of compound 2.01described above. Method 2 was followed for the synthetic sequence,wherein butyryl chloride was used in step a instead of propionylchloride, 4-ethoxyaniline was used in step b instead of 4-fluoroaniline,and 2-ethoxy-1-aminoethane was used in step d instead of1-(2-aminoethyl)morpholine. Characterization data for compound 2.11follows: colorless, viscous oil. ¹H NMR (d₆-DMSO; T=140° C.) δ0.80 (t,3H, J=7.6 Hz), 0.94 (t, 3H, J=6.8 Hz), 1.35 (t, 3H, J=6.8 Hz), 1.59-1.70(m, 1H), 2.20-2.30 (m, 1H), 2.77 (s, 3H), 3.22-3.42 (m, 4H), 3.47-3.65(m, 2H), 4.10 (q, 2H, J=6.8 Hz), 5.01 (br q, 1H), 6.98-7.12 (m, 2H),7.15-7.27 (m, 4H), 7.29-7.36 (m, 2H), 7.41-7.47 (m, 2H), 7.51-7.56 (m,3H), 7.59-7.63 (m, 2H), 7.67 (dd, 1H, J₁=7.6 Hz, J₂=7.8 Hz) ppm. At roomtemperature, compound exists as a mixture of cis/trans amide rotamers,ca. 5:3 by ¹H NMR (CDCl₃; T=25° C.) δ_(major) 4.65 (dd, 1.7H, J₁=4.8 Hz,J₂=10.0 Hz) and δ_(minor) 5.39 (dd, 1.0H, J₁=J₂=7.2 Hz) ppm. MS (ESI⁺)604.2 [MH]⁺

Synthesis of Compound 2.12

Compound 2.12 was prepared following the synthesis of compound 2.01described above. Method 2 was followed for the synthetic sequence,wherein 2-aminonicotinic acid was used in step a instead of2-amino-6-methylbenzoic acid, 4-ethoxyaniline was used in step b insteadof 4-fluoroaniline, 2-ethoxy-1-aminoethane was used in step d instead of1-(2-aminoethyl)morpholine, and 4-trifluoromethylphenylacetic acid wasused in step e instead of biphenylacetyl chloride. Characterization datafor compound 2.12 follows: colorless, viscous oil. ¹H NMR (d₆-DMSO;T=140° C.) δ0.96 (t, 3H, J=7.2 Hz), 1.36 (t, 3H, J=7.2 Hz), 1.47 (d, 3H,J=6.4 Hz), 3.29-3.40 (m, 2H), 3.42-3.51 (m, 2H), 3.54-3.64 (m, 2H); 4.11(q, 2H, J=6.8 Hz), 5.20 (q, 1H, J=7.2 Hz), 7.00-7.10 (m, 2H), 7.23-7.41(m, 4H), 7.53-7.60 (m, 3H), 8.50 (dd, 1H, J₁=2.4 Hz, J₂=8.4 Hz), 9.00(dd, 1H, J₁=2.0 Hz, J₂=4.4 Hz) ppm. At room temperature, compound existsas a mixture of cis/trans amide rotamers, ca. 3:2 by ¹H NMR (CDCl₃;T=25° C.) δ_(minor) 5.00 (q, 1.0H, J=6.0 Hz) and δ_(major) 5.38 (q,1.4H, J=7.2 Hz) ppm. MS (ESI⁺) 569.3 [MH]⁺.

Example 3 Synthesis of 3.01

The synthesis of compound 3.01 in five steps from commercially availablestarting materials provides an example of a 3H-quinazolin-4-onesynthesis in enantiomerically enriched form. Scheme 3 provides anoverview of the synthetic route, for which the experimental detailsfollow.

(R)-2-(1-N-BOC-aminoethyl)-3-(4-ethoxyphenyl)-3H-quinazoline-4-one(XII). To a solution of anthranilic acid (411 mg, 3.0 mmol, 1.0 equiv)and N-BOC-D-alanine (568 mg, 3.0 mmol, 1.0 equiv) in 3.0 mL of anhydrouspynidine was added 0.96 mL of triphenylphosphite (1.14 g, 3.6 mmol, 1.2equiv) at room temperature. The resulting yellow solution was stirred at50° C. for 20 h. p-Phenetidine (453 mg, 3.3 mmol, 1.1 equiv) was addedvia syringe. The reaction mixture was stirred for another 2 h at 50° C.,cooled to room temperature, and evaporated in vacuo to remove most ofpyridine. The residue in 15 mL of diethyl ether was washed successivelytwice with 9 mL of 5% aqueous phosphoric acid, twice with 9 mL of 1 MNaOH, once with 5 mL of pH 7 phosphate buffer (0.5 M KH₂PO₄ and 0.5 MK₂HPO₄), and once with 9 mL of brine. The organic layer was dried overNa₂SO₄ and evaporated in vacuo to give a brown residue, which wasrecrystallized from a mixture of 3 mL of EtOAc and 12 mL of heptane togive 0.51 g of compound XII as a white solid. The mother liquor wasconcentrated in vacuo to give a brown residue, which was recrystallizedfrom a mixture of 1 mL of EtOAc and 4 mL of heptane to give a secondcrop of 0.13 g of XII as a light yellow solid. m.p. 143.7° C. ¹H NMR(DMSO-d₆) δ1.19 (d, J=6.4 Hz, 3H), 1.32 (s, 9H), 1.37 (t, J=6.8 Hz, 3H),4.10 (q, J=6.9 Hz, 2H), 4.24 (m, 1H), 7.09 (m, 2H), 7.28 (d, J=8.0 Hz,2H), 7.39 (dd, J=8.4, 2.0 Hz, 1H), 7.54 (t, J=7.6 Hz, 1H), 7.69 (d,J=8.0 Hz, 1H), 7.85 (t, J=8.0 Hz, 1H), 8.11 (d, J=8.0 Hz, 1H) ppm. MS(ESI⁺) m/z 410.2 [M+H]⁺.

(R)-2-(1-Aminoethyl)-3-(4-ethoxyphenyl)-3H-quinazoline-4-one (XIII). Toa suspension of XII (9.39 g, 22.9 mmol, 1.0 equiv) in 45 mL of anhydrousacetonitrile was added 3.43 mL of iodotrimethylsilane (4.82 g, 24.1mmol, 1.05 equiv) dropwise via syringe over 15 min. After stirring foranother 45 min at room temperature, all starting material XII had beenconsumed. The resulting mixture was partitioned between 50 mL of 1 MNH₄OH and 90 mL of ether. The aqueous layer was extracted two more timeswith 30 mL of ether. The combined ether extract was washed once with 40mL of brine. The organic layer was dried over Na₂SO₄ and evaporated invacuo to give a light gray solid. Recrystallization of this crudeproduct from 25 mL of dioxane gave 4.2 g of XIII as a white solid. Themother liquor was concentrated in vacuo to give a light gray solid whichwas triturated with 15 mL of ether to give 1.8 g of additional productas a off-white solid. Total yield was 6.0 g. m.p. 179.9° C. ¹H NMR(DMSO-d₆) δ1.16 (d, J=6.4 Hz, 3H), 1.38 (t, J=7.0 Hz, 3H), 2.25 (br s,2H), 3.51 (q, J=6.4 Hz, 1H), 4.11 (q, J=6.9 Hz, 2H), 7.08 (m, 2H), 7.36(m, 2H), 7.52 (t, J=7.6 Hz, 1H), 7.70 (d, J=8.0 Hz, 1H), 7.85 (t, J=7.8Hz, 1H), 8.11 (d, J=8.0 Hz, 1H) ppm. MS (ESI⁺) m/z 310.1 [M+H]⁺.

(R)-2-((N-3-Picolyl)-N-(4-trifluoromethylphenylacetyl)-1-aminoethyl)-3-(4-ethoxyphenyl)-3H-quinazoline-4-one (3.01). 3-Picolylchloridehydrochloride (4.27 g, 26 mmol, 1.15 equiv), KI (4.32 g, 26.0 mmol, 1.15equiv), and 60 mL of DMPU were mixed in a 200 mL flask. The mixture wasvigorously stirred for 1 h at room temperature. To the resulting yellowmixture was added compound XIII (7.0 g, 22.6 mmol, 1.0 equiv) and K₂CO₃(9.38 g, 67.9 mmol, 3.0 equiv). The mixture was stirred at roomtemperature for 14 h. Additional 3-picolylchloride hydrochloride (740mg, 4.51 mmol, 0.2 equiv) was added and the mixture was stirred foranother 8 h at room temperature.

To the above reaction mixture was added 4-trifluoromethylphenylaceticacid (5.08 g, 24.9 mmol, 1.1 equiv), HOBT (4.58 g, 33.9 mmol, 1.5equiv), and 20 mL of dichloromethane at room temperature. EDC (13.0 g,67.8 mmol, 3.0 equiv) was then added portionwise over 15 min. After theinitial gas evolution had subsided, the mixture was stirred vigorouslyat room temperature for another 14 h. The reaction mixture was pouredinto a mixture of 180 mL of 10% citric acid and 150 mL of ether. Theaqueous layer was extracted twice with 100 mL of ether. The combinedether extract was washed twice with 60 mL of 2% citric acid, twice with50 mL of saturated NaHCO₃, and once with 100 mL of brine. The organiclayer was dried over Na₂SO₄ and evaporated in vacuo to give a orangefoam, which was recrystallized from 20 mL of 1:1 heptane/i-PrOH to give6.50 g of compound 3.01 as a light yellow solid. m.p. 176.3° C. ¹H NMR(DMSO-d₆, T=140° C.) δ1.36 (t, J=6.9 Hz, 3H), 1.41 (d, J=6.9 Hz, 3H),1.53 (d, J=19.6 Hz, 1H), 3.18 (br, 1H), 4.12 (q, J=6.9 Hz, 2H), 4.70 (d,J=16.7 Hz, 1H), 4.76 (d, J=16.6 Hz, 1H), 5.28 (q, J=6.6 Hz, 1H), 7.08(br, 3H), 7.15 (dd, J=7.7, 4.8 Hz, 1H), 7.27 (d, J=8.0 Hz, 2H), 7.37(br, 1H), 7.48-7.58 (m, 4H), 7.68 (d, J=7.7 Hz, 1H), 7.85 (m, 1H), 8.10(m, 1H), 8.34 (d, J=4.5 Hz, 1H), 8.37 (s, 1H) ppm. At room temperature,this compound exists as a mixture of cis/trans amide rotamers, ca.1.83:1 by ¹H NMR (DMSO-d₆, T=25° C.) δ5.11 (q, J=6.8 Hz, 1H) & 5.28 (q,J=6.8 Hz, 1H) ppm. MS (ESI⁺) m/z 587.3 [M+H]⁺.

(R)-2-((N-3-Picolyl)-N-(4-trifluoromethylphenylacetyl)-1-aminoethyl)-3-(4-ethoxyphenyl)-3H-quinazoline-4-one hydrochloride (3.01.HCl). To asolution of compound 3.01 (3.55 g, 6.05 mmol, 1.0 equiv) in 25 mL ofether and 25 mL of dichloromethane was added a 1.0 M solution of HCl inether (12.1 mL, 12.1 mmol, 2.0 equiv) dropwise via syringe, followed byanother 50 mL of ether. The resulting suspension was stirred at roomtemperature for 1 h. The precipitates were collected by filtration. Thesolids were washed twice with 30 mL of ether and air dried in the darkto give 3.74 g of the product as a white powder. m.p. 186.2° C. At roomtemperature, this compound exists as a mixture of cis/trans amiderotamers, ca. 1.78:1 by ¹H NMR (DMSO-d₆, T=25° C.) δ1.48 (d, J=6.4 Hz,3H) & 1.22 (d, J=7.2 Hz, 3H) ppm. At 140° C., the ¹H NMR spectra of3.01.HCl was identical to that of 3.01. MS (ESI⁺) m/z 587.3 [M+H]⁺.Chiral HPLC showed the enantiomeric ratio of this product to be 98:2R/S.

Synthesis of Compound 3.02

The synthesis of compound 3.02 in five steps from commercially availablestarting materials provides an example of a 3H-quinazolin-4-onesynthesis in racemic form. Scheme 4 provides an overview of thesynthetic route, for which the experimental details follow.

2-(1-N-BOC-aminoethyl)-3-(4-fluorophenyl)-3H-quinazoline-4-one (XIV). Toa solution of anthranilic acid (2.74 g, 20 mmol, 1.0 equiv) andN-BOC-D-alanine (3.78 g, 20 mmol, 1.0 equiv) in 20 mL of anhydrouspyridine was added 5.24 mL of triphenylphosphite (6.21 g, 20 mmol, 1.0equiv) at room temperature. The resulting yellow solution was stirred at100° C. for 4 h. 4-Fluoroaniline (2.22 g, 20 mmol, 1.0 equiv) was addedvia syringe. The reaction mixture was stirred for another 3 h at 100°C., cooled to room temperature, and evaporated in vacuo to give a brownresidue. This residue was dissolved in 50 mL of EtOAc. The mixture waswashed successively twice with 40 mL of 5% aqueous phosphoric acid, oncewith 20 mL of saturated NaHCO₃, and once with 40 mL of brine. Theorganic layer was dried over Na₂SO₄ and evaporated in vacuo to give abrown residue, which was purified by silica gel chromatography to give2.40 g of compound XIV as a light yellow solid. ¹H NMR (DMSO-d₆) δ1.22(d, J=6.8 Hz, 3H), 1.31 (s, 9H), 4.21 (m, 1H), 7.30 (m, 1H), 7.42 (m,3H), 7.58 (m, 2H), 7.71 (d, J=8.0 Hz, 1H), 7.88 (t, J=7.8 Hz, 1H), 8.11(d, J=8.0 Hz, 1H) ppm. MS (ESI⁺) m/z 384.0 [M+H]⁺.

2-(1-Aminoethyl)-3-(4-fluorophenyl)-3H-quinazoline-4-one hydrochloride(XV). To a solution of compound XIV (2.30 g, 6.0 mmol, 1.0 equiv) in 6.0mL of EtOAc was added 6.0 mL of a 4.0 M solution of HCl in dioxane (24mmol, 4.0 equiv) at room temperature. After the resulting solution wasstirred at room temperature for 1.5 h, it was evaporated in vacuo togive a light gray solid. This crude product was dissolved in 9 mL ofdichloromethane. To this stirring solution was added a total of 36 mL ofether via an additional funnel. The precipitates were collected byvacuum filtration, washed twice with 10 mL of ether, and air-dried togive 1.1 g of compound XV as a slightly off-white solid. ¹H NMR(DMSO-d₆) δ1.31 (d, J=6.8 Hz, 3H), 3.89 (m, 1H), 7.20 (t, J=4.8 Hz, 1H),7.48 (m, 2H), 7.67 (m, 2H), 7.77 (d, J=8.0 Hz, 1H), 7.95 (t, J=7.6 Hz,1H), 8.18 (d, J=8.0 Hz, 1H), 8.53 (br, 3H) ppm. MS (ESI⁺) m/z 284.0[M+H]⁺.

2-((N-3-Picolyl)-N-(4-trifluoromethylphenylacetyl)-1-aminoethyl)-3-(4-fluorophenyl)-3H-quinazoline-4-one(3.02). 3-Picolylchloride hydrochloride (333 mg, 2.03 mmol, 1.15 equiv),KI (59 mg, 0.35 mmol, 0.20 equiv), compound XV (0.56 g, 1.77 mmol, 1.0equiv), and K₂CO₃ (513 mg, 3.71 mmol, 2.1 equiv) were added to 2.5 mL ofDMF. The mixture was vigorously stirred for 14 h at room temperature.The mixture was poured into 20 mL of 10% Na₂CO₃ and extracted four timeswith 10 mL of EtOAc. The combined EtOAc extract was washed once with 20mL of brine, dried over Na₂SO₄, and evaporated in vacuo to give anorange colored foam, which was used without further purification.

To the above crude product was added 4-trifluoromethylphenylacetic acid(542 mg, 2.66 mmol, 1.5 equiv), EDC (594 mg, 3.10 mmol, 1.75 equiv) HOBT(419 mg, 3.00 mmol, 1.7 equiv), N-methylmorpholine (304 mg, 3.00 mmol,1.7 equiv), and 6.0 mL of dichloromethane at room temperature. Themixture was stirred at room temperature for 14 h. The reaction mixturewas poured into a 20 mL of 10% citric acid, and extracted twice with 15mL of EtOAc. The combined EtOAc extract was washed once with 20 mL ofsaturated NaHCO₃, and once with 20 mL of brine. The organic layer wasdried over Na₂SO₄ and evaporated in vacuo to give a brown residue, whichwas purified by silica gel chromatography to give 169 mg of compound3.02 as a white solid. m.p. 167.0° C. ¹H NMR (DMSO-d₆, T=140° C.) δ1.40(d, J=6.7 Hz, 3H), 3.32 (br, 1H), 3.59 (d, J=15.8 Hz, 1H), 4.73 (d,J=17.6 Hz, 1H), 4.81 (d, J=17.2 Hz, 1H), 5.26 (q, J=6.5 Hz, 1H), 7.18(dd, J=7.7 Hz, 4.8 Hz, 1H), 7.29 (d, J=8.1 Hz, 2H), 7.34 (m, 3H), 7.55(m, 5H), 7.67 (d, J=8.0 Hz, 1H), 7.86 (dd, J=7.6, 1.6 Hz, 1H), 8.11 (d,J=8.0 Hz, 1H), 8.36 (q, J=4.8 Hz, 1H), 8.40 (s, 1H) ppm. At roomtemperature, this compound exists as a mixture of cis/trans amiderotamers, ca. 0.96:1 molar ratio (DMSO-d₆, T=25° C.) d 5.10 (q, J=6.8Hz, 1H) & 5.31 (q, J=6.8 Hz, 1H) ppm. MS (ESI⁺) m/z 561.2 [M+H]⁺. ChiralHPLC showed the enantiomeric ratio of this product to be ca. 1:1 R/S.

2-((N-3-Picolyl)-N-(4-trifluoromethylphenylacetyl)-1-aminoethyl)-3-(4-fluorophenyl)-3H-quinazoline-4-one hydrochloride (3.02.HCl). To a solution ofcompound 3.02 (50 mg, 89 mmol, 1.0 equiv) in 2 mL of dichloromethane wasadded a 1.0 M solution of HCl in ether (180 mL, 0.18 mmol, 2.0 equiv)dropwise via syringe, followed by another 5 mL of ether. The resultingsuspension was stirred at room temperature for 1 h. The precipitateswere collected by filtration. The solids were washed twice with 30 mL ofether and air dried in the dark to give 47 mg of the product as a whitepowder. mp 122.7° C. At room temperature, this compound exists as amixture of cis/trans amide rotamers, ca. 0.93:1 by ¹H NMR (DMSO-d₆,T=25° C.) δ5.05 (q, J=6.8 Hz, 1H) & 5.18 (q, J=6.8 Hz, 1H) ppm. MS(ESI⁺) m/z 561.2 [M+H]⁺.

Synthesis of Compound 3.03 and Compound 3.03.HCl

The synthesis of compound 3.03 followed the method described forcompound 3.02. 2-Picolylchloride hydrochloride was used in place of3-picolylchloride hydrochloride in step 3 of the synthetic sequence.Characterization of the products follows.

2-((N-(2-Picolyl)-N-(4-trifluoromethylphenylacetyl)-1-aminoethyl)-3-(4-fluorophenyl)-3H-quinazoline-4-one(3.03) was obtained as a white solid from compound XV. Mp 159.2° C. Atroom temperature, this compound exists as a mixture of cis/trans amiderotamers, ca. 0.23:1 by ¹H NMR (DMSO-d₆, T=25° C.) δ5.05 (q, J=6.8 Hz,1H) & 5.36 (q, J=6.8 Hz, 1H) ppm. MS (ESI⁺) m/z 561.2 [M+H]⁺.

2-((N-2-Picolyl)-N-(4-trifluoromethylphenylacetyl)-1-aminoethyl)-3-(4-fluorophenyl)-3H-quinazoline-4-one hydrochloride (3.03.HCl) was obtained as a whitesolid from compound 3.03. At room temperature, this compound exists as amixture of cis/trans amide rotamers, ca. 0.64:1 by ¹H NMR (DMSO-d₆,T=25° C.) δ5.04 (q, J=6.8 Hz, 1H) & 5.35 (q, J=6.8 Hz, 1H) ppm. MS(ESI⁺) m/z 561.2 [M+H]⁺.

Synthesis of Compounds 3.04 and 3.04.HCl

The synthesis of compound 3.04 followed the method described forcompound 3.02. p-Phenetidine was used in place of 4-fluoroaniline instep 1 of the synthetic sequence. 2-Picolylchloride hydrochloride wasused in place of 3-picolylchloride hydrochloride in step 3 of thesynthetic sequence. Characterization of the products follows.

2-((N-2-Picolyl)-N-(4-trifluoromethylphenylacetyl)-1-aminoethyl)-3-(4-ethoxyphenyl)-3H-quinazoline-4-one (3.04) was obtained as a white solid from thehydrochloride salt of racemic compound XIII. Mp 167.5° C. ¹H NMR(DMSO-d₆, T=140° C.) δ1.35 (d, J=6.9 Hz, 3H), 1.37 (t, J=7.4 Hz, 3H),3.49 (br, 1H), 3.64 (m, 1H), 4.10 (q, J=9.1 Hz, 2H), 4.78 (d, J=17.2 Hz,1H), 4.84 (d, J=17.2 Hz, 1H), 5.38 (q, J=6.2 Hz, 1H), 7.02 (br, 2H),7.09 (dd, J=6.7, 4.8 Hz, 1H), 7.18 (d, J=7.9 Hz, 1H), 7.30 (m, 4H),7.46-7.58 m, 4H), 7.61 (d, J=8.3 Hz, 1H), 7.79 (m, 1H), 8.07 (dd, J=8.0,1.4 Hz, 1H), 8.36 (m, 1H) ppm. At room temperature, this compound existsas a mixture of cis/trans amide rotamers, ca. 0.42:1 molar ratio(DMSO-d6, T=25° C.) d 5.12 (q, J=6.8 Hz, 1H) & 5.34 (q, J=6.8 Hz, 1H)ppm. MS (ESI⁺) m/z 587.2 [M+H]⁺.

2-((N-(2-Picolyl)-N-(4-trifluoromethylphenylacetyl)-1-aminoethyl)-3-(4-ethoxyphenyl)-3H-quinazoline-4-onehydrochloride (3.04.HCl) was obtained as a white solid from compound3.04. Mp 162.6° C. At room temperature, this compound exists as amixture of cis/trans amide rotamers, ca. 1.45:1 molar ratio (DMSO-d6,T=25° C.) d 1.51 (d, J=6.4 Hz, 1H) & 1.24 (d, J=7.2 Hz, 1H) ppm. MS(ESI) m/z 587.2 [M+H]⁺.

Synthesis of Compound 3.05

The synthesis of compound 3.05 is closely related to that of compound3.02 described above. Scheme 5 provides an overview of synthetic route.Compound 3.05 also served as a common precursor for a series of closelyrelated compounds.

2-(1-Aminoethyl)-3-(4-iodophenyl)-3H-quinazoline-4-one hydrochloride(XVI) The product was a white solid. ¹H NMR (DMSO-d₆) δ1.31 (d, J=6.8Hz, 3H), 3.89 (m, 1H), 7.31 (d, J=7.6 Hz, 1H), 7.42 (d, J=7.2 Hz, 1H),7.63 (t, J=7.6 Hz, 1H), 7.76 (d, J=8.0 Hz, 1H), 7.97 (m, 3H), 8.14 (d,J=8.0 Hz, 1H), 8.51 (br, 3H) ppm. MS (ESI⁺) m/z 392.0 [M+H]⁺.

2-((N-2-Ethoxyethyl)-N-(4-trifluoromethylphenylacetyl)-1-aminoethyl)-3-(4-iodophenyl)-3H-quinazoline-4-one (3.05) was obtained as a whitesolid from compound XVI. Mp 181.8° C. At room temperature, this compoundexists as a mixture of cis/trans amide rotamers, ca. 0.64:1 by ¹H NMR(DMSO-d₆, T=25° C.) δ4.89 (q, J=6.0 Hz, 1H) & 5.22 (q, J=6.4 Hz, 1H)ppm. MS (ESI⁺) m/z 650.2 [M+H]⁺.

2-((N-2-Ethoxyethyl)-N-(4-trifluoromethylphenylacetyl)-1-aminoethyl)-3-(4-cyanophenyl)-3H-quinazoline-4-one (3.06). 3.05 (150 mg, 0.23 mmol,1.0 equiv) was dissolved in 0.5 mL of anhydrous DMF. CuCN (31 mg, 0.35mmol, 1.5 equiv) was added. The resulting mixture was heated to 130° C.for 16 h. The mixture was cooled to room temperature and diluted with 15mL of EtOAc. The mixture was filtered through a short column of silicagel, which was further eluted with 50 mL of EtOAc. The eluent wasconcentrated in vacuo to give a yellow residue, which was purified bypreparative TLC to give 95 mg of compound 3.06 as a white solid. Mp197.0° C. ¹H NMR (DMSO-d₆, T=140° C.) δ0.98 (t, J=6.9 Hz, 3H), 1.44 (d,J=6.8 Hz, 3H), 3.30-3.65 m, 8H), 5.16 (q, J=6.2 Hz, 1H), 7.33 (d, J=8.0Hz, 2H), 7.50-7.77 (m, 6H), 7.72-7.95 (m, 3H), 8.15 (dd, J=7.9, 1.5 Hz,1H) ppm. At room temperature, this compound exists as a mixture ofcis/trans amide rotamers, ca. 0.64:1 molar ratio in DMSO. ¹H NMR(DMSO-d₆, T=25° C.) δ4.84 (q, J=6.4 Hz, 1H) & 5.22 (q, J=6.4 Hz, 1H)ppm. MS (ESI⁺) m/z 650.2 [M+H]⁺.

2-((N-2-Ethoxyethyl)-N-(4-trifluoromethylphenylacetyl)-1-aminoethyl)-3-(4-trimethylsilyIethynylphenyl)-3H-quinazoline-4-one (3.07). Compound3.05 (150 mg, 0.23 mmol, 1.0 equiv) was dissolved in 1.0 mL of anhydrousTHF. Trimethylsilylacetylene (45 mg, 0.46 mmol, 2.0 equiv), CuI (87 mg,0.46 mmol, 2.0 equiv), Pd(PPh₃)₂Cl₂ (32 mg, 0.046 mmol, 0.20 equiv), andtriethylamine (92 mg, 0.91 mmol, 4.0 equiv) were added sequentially. Theresulting mixture was heated to 50° C. for 3 h. The mixture was cooledto room temperature and diluted with 15 mL of EtOAc. The mixture wasfiltered through a short column of silica gel, which was further elutedwith 50 mL of EtOAc. The eluent was concentrated in vacuo to give ayellow residue, which was purified by preparative TLC to give 105 mg3.07 as a white solid. Mp 185.3° C. At room temperature, this compoundexists as a mixture of cis/trans amide rotamers, ca. 0.54:1 molar ratioin DMSO. ¹H NMR (DMSO-d₆, T=25° C.) δ4.88 (q, J=6.8 Hz, 1H) & 5.25 (q,J=6.8 Hz, 1H) ppm. MS (ESI⁺) m/z 620.2 [M+H]⁺.

2-((N-2-Ethoxyethyl)-N-(4-trifluoromethylphenylacetyl)-1-aminoethyl)-3-(4-ethynylphenyl)-3H-quinazoline-4-one (3.08). Compound 3.07 (57 mg,92 μmol, 1.0 equiv) was dissolved in 1.0 mL of anhydrous THF. A 1.0 Msolution of tetrabutylammonium fluoride in THF (101 μL, 0.101 mmol, 1.1equiv) was added at room temperature. The resulting mixture was stirredat room temperature for 15 min. To the reaction mixture was added 100 μLof saturated aqueous NH₄Cl and 15 mL of EtOAc. After stirring at roomtemperature for another 15 min, the mixture was dried over Na2SO4, andfiltered through a short column of silica gel, which was further elutedwith 50 mL of EtOAc. The eluent was concentrated in vacuo to give ayellow residue, which was purified by preparative TLC to give 39 mg ofcompound 3.08 as a white solid. Mp 186.7° C. At room temperature, thiscompound exists as a mixture of cis/trans amide rotamers, ca. 0.54:1molar ratio in DMSO. ¹H NMR (DMSO-d₆, T=25° C.) δ4.87 (q, J=6.0 Hz, 1H)& 5.20 (q, J=6.8 Hz, 1H) ppm. MS (ESI⁺) m/z 548.2 [M+H]⁺.

Synthesis of Compounds 3.09 and 3.10

Compounds 3.09 and 3.10 were synthesized in the same manner as compound3.07 (See Scheme 7). 3,3-Dimethyl-1-butyne, and 2-methyl-3-butyn-2-olwere used respectively, instead of trimethylsilylacetylene.Characterization of the products follows.

2-((N-2-Ethoxyethyl)-N-(4-trifluoromethylphenylacetyl)-1-aminoethyl)-3-(4-(t-butylethynyl)phenyl)-3H-quinazoline-4-one (3.09) was obtainedfrom compound 3.05 in as a white solid. Mp 189.9° C. At roomtemperature, this compound exists as a mixture of cis/trans amiderotamers, ca. 0.69:1 molar ratio in DMSO. ¹H NMR (DMSO-d₆, T=25° C.)δ4.89 (q, J=6.4 Hz, 1H) & 5.25 (q, J=6.4 Hz, 1H) ppm. MS (ESI⁺) m/z604.2 [M+H]⁺.

2-((N-2-Ethoxyethyl)-N-(4-trifluoromethylphenylacetyl)-1-aminoethyl)-3-(4-(3-hydroxy-3-methyl-1-butynyl)phenyl)-3H-quinazoline-4-one (3.10)was obtained from compound 3.05 as a white solid. Mp 162.2° C. At roomtemperature, this compound exists as a mixture of cis/trans amiderotamers, ca. 0.69:1 molar ratio in DMSO. ¹H NMR (DMSO-d₆, T=25° C.)δ4.89 (q, J=6.8 Hz, 1H) & 5.24 (q, J=6.4 Hz, 1H) ppm. MS (ESI⁺) m/z606.3 [M+H]⁺.

Synthesis of Compounds 3.11 and 3.12

Compounds 3.11 and 3.12 were synthesized in the same manner as compounds3.07 and 3.08 (Scheme 7). Compound 1.42 was used as starting material inboth cases, instead of compound 3.05. Characterization of the productsfollows.

2-((N-2-Methoxyethyl)-N-(4-phenylphenylacetyl)-1-aminoethyl)-3-(4-trimethylsilylethynyl)phenyl)-3H-quinazoline-4-one (3.11) was obtained from compound 1.42 as a whitesolid. At room temperature, this compound exists as a mixture ofcis/trans amide rotamers, ca. 0.61:1 molar ratio in DMSO. ¹H NMR(DMSO-d₆, T=25° C.) δ4.91 (q, J=6.4 Hz, 1H) & 5.21 (q, J=6.4 Hz, 1H)ppm. MS (ESI⁺) m/z 614.3 [M+H]⁺.

2-((N-2-Methoxyethyl)-N-(4-phenylphenylacetyl)-1-aminoethyl)-3-(4-ethynylphenyl)-3H-quinazoline-4-one (3.12) was obtained from compound 1.42 as a whitesolid. Mp 73.3° C. At room temperature, this compound exists as amixture of cis/trans amide rotamers, ca. 0.67:1 molar ratio in DMSO. ¹HNMR (DMSO-d₆, T=25° C.) δ4.92 (q, J=6.4 Hz, 1H) & 5.17 (q, J=6.8 Hz, 1H)ppm. MS (ESI⁺) m/z 542.2 [M+H]⁺.

Synthesis of Compounds 3.13 and 3.14. Compounds 3.13 and 3.14 weresynthesized in the same reaction from compound 3.05. Experimentaldetails follow.

2-((N-2-Ethoxyethyl)-N-(4-trifluoromethylphenylacetyl)-1-aminoethyl)-3-phenyl-3H-quinazoline-4-one (3.13) and2-((N-2-Ethoxyethyl)-N-(4-trifluoromethylphenylacetyl)-1-aminoethyl)-3-(4-ethylphenyl)-3H-quinazoline-4-one (3.14) To asolution of compound 3.05 (98 mg, 0.15 mmol, 1.0 equiv) and Pd(PPh₃)₄(35 mg, 30 μmol, 0.20 equiv) in 1.0 mL of THF was added Et₂Zn (37 mg,0.30 mmol, 2.0 equiv) via syringe. The darkened reaction mixture wasstirred at 50° C. for 3 h. After cooling to room temperature, thereaction mixture was diluted with 20 mL of EtOAc, and washedsuccessively with 10 mL of 1 M HCl, 10 mL of saturated NaHCO₃, and 10 mLof brine. The organic layer was dried over Na2SO4, and evaporated invacuo to give a brown residue, which was purified by preparative TLC togive 18 mg of compound 3.13 and 27 mg of compound 3.14. Both are whitesolids. Characterization of these two products follows.

Compound 3.13. At room temperature, this compound exists as a mixture ofcis/trans amide rotamers, ca. 0.69:1 molar ratio in CDCl₃. ¹H NMR(CDCl₃, T=25° C.) δ0.71 (d, J=7.0 Hz, 3H) & 1.15 (d, J=7.0 Hz, 1H) ppm.MS (ESI^(+) m/z) 524.3 [M+H]⁺.

Compound 3.14. At room temperature, this compound exists as a mixture ofcis/trans amide rotamers, ca. 0.69:1 molar ratio in CDCl₃. ¹H NMR(CDCl₃, T=25° C.) δ4.88 (q, J=6.8 Hz, 1H) & 5.34 (q, J=6.8 Hz, 1H) ppm.MS (ESI⁺) m/z 552.2 [M+H]⁺.

Synthesis of Compound 3.15

Compound 3.15 was synthesized from compound1.42 using Pd catalyzedhydrogenation. Experimental details follow.

2-((N-2-Methoxyethyl)-N-(4-phenylphenylacetyl)-1-aminoethyl)-3-phenyl-3H-quinazoline-4-one (3.15) To a solution of compound 1.42 (25 mg, 39μmol, 1.0 equiv) in a mixture of 1.0 mL of MeOH and 1.0 mL ofdichloromethane, was added 10% Pd on carbon (83 mg, 78 μmol, 2.0 equiv).Excess hydrogen was introduced using a balloon. After stirring at roomtemperature for 2 h. The reaction mixture was diluted with 5 mL ofdichloromethane, and filtered through a pad of Celite. The filtrate wasconcentrated in vacuo to give a yellow oil, which was passed through ashort column of silica gel, eluted with EtOAc. The eluent wasconcentrated in vacuo to give 17 mg of compound 3.15 as a colorless oil.At room temperature, this compound exists as a mixture of cis/transamide rotamers, ca. 1.06:1 molar ratio in DMSO. ¹H NMR (DMSO-d₆, T=25°C.) δ4.94 (q, J=6.4 Hz, 1H) & 5.08 (q, J=6.8 Hz, 1H) ppm. MS (ESI⁺) m/z518.3 [M+H]⁺.

Synthesis of Compound 3.16

Compound 3.16 was synthesized from the racemic form of compound XII asdescribed in the experimental details below.2-((N-3-Picolyl)-N-(t-butoxycarbonyl)-1-aminoethyl)-3-(4-ethoxyphenyl)-3H-quinazoline-4-one (3.16). To a solution of racemic compound XII (124mg, 0.30 mmol, 1.0 equiv) in 0.60 mL of DMF, was added 3-picolylchioridehydrochloride (55 mg, 0.33 mmol, 1.1 equiv), KI (50 mg, 0.30 mmol, 1.0equiv), and NaH (60% suspension in mineral oil, 25 mg, 0.62 mmol, 2.05equiv). After stirred at room temperature for 16 h, the reaction mixturewas poured into 10 mL of 5% aqueous H₃PO₄. The resulting mixture wasextracted twice with 10 mL of EtOAc. The organic layer was washed with10 mL of NaHCO₃ and 10 mL of brine, dried over Na₂SO₄ and concentratedin vacuo to give a yellow oil, which was purified by preparative TLC togive 33 mg of compound 3.16 as a white solid. Mp 67.5° C. At roomtemperature, this compound exists as a mixture of cis/trans amiderotamers, ca. 1.11:1 molar ratio in DMSO. ¹H NMR (DMSO-d₆, T=25° C.)δ5.03 (m, 1H) & 5.12 (m, 1H) ppm. MS (ESI⁺) m/z 510.3 [M+H]⁺.

Synthesis of Compound 3.16a

(1-N-BOC-aminoethyl)-3-(4-ethoxyphenyl)-2-{(1R)-1-[(pyridin-3-ylmethyl)-amino]-ethyl}-3H-pyrido[2,3-d]pyrimidin-4-one(XVII precursor). To a 3 L round bottom flask equipped with additionfunnel, mechanical stirrer and temperature probe was added 102.60 g(542.26 mmol) of N-(tert-butoxycarbonyl)-D-alanine in 1.2 L ofdichloromethane (DCM) under a nitrogen atmosphere. The solution wascooled to −20° C. and 150.00 ml (1364.31 mmol) of N-methyl morpholineadded followed by the addition of a solution containing 140.1 ml (1084mmol) of iso-butylchloroformate in 360 ml of DCM over 40 min. whilemaintaining the reaction temperature below −20° C. After completeaddition, the reaction was allowed to stir for 45 min. and 75.00 g(542.97 mmol) of 2-aminonicotinic acid added. The reaction was allowedto warm to room temperature overnight. The reaction was diluted with 1.5L DCM and washed with 1.0 N hydrochloric acid (2×750 ml) and brine(1×500 ml). The organic phase was dried over magnesium sulfate,filtered, and concentrated in vacuo to give 175.0 g of a yellow-orangeoil. The material was used without further purification in the nextstep.

A solution containing the crude material from above dissolved in 2 L DCMwas cooled to 20° C. under a nitrogen atmosphere and 69.00 ml (535.68mmol) of p-phenetidine was added over 5 minutes. After stirring withgradual warming to 0° C. the reaction mixture was transferred to aseparatory funnel and washed with 1.0 N hydrochloric acid (2×500 ml),saturated sodium bicarbonate solution (2×1 L), and brine (1×1 L). Theorganic phase was dried over magnesium sulfate, filtered, andconcentrated in vacuo to give 175.2 g of crude bis-amide. The materialwas used without further purification in the next step.

A solution containing the crude bis-amide prepared above in 2.3 L of DCMand 50.0 ml (454.7 mmol) of N-methyl morpholine was cooled to −20° C.under a nitrogen atmosphere and 53.0 ml (408.6 mmol) ofiso-butylchloroformate was added dropwise over a period of 5 minutes.Upon completed addition of the chloroformate HPLC analysis indicated nobis-amide remained. The reaction mixture was transferred to a separatoryfunnel and washed with 1.0 N hydrochloric acid (2×1 L), saturatedbicarbonate solution (1×1 L), and brine (1×1 L). The organic phase wasdried over magnesium sulfate, filtered, and concentrated in vacuo togive 205 g of a brown, viscous oil. This product was dissolved in 500 mlof methyl tert-butyl ether and allowed to stir until the product beganto precipitate from the solution. Heptane was then added (1000 ml) andstirring continued. The resulting precipitate was collected byfiltration, washed with heptane, and dried to afford 78.9 g of productas an off-white solid. ¹H NMR (CDCl₃) δ8.99 (dd, J₁=2.0 Hz, J₂=4.4 Hz,1H), 8.60 (dd, J₁=2.0 Hz, J₂=8.0 Hz, 1H), 7.44 (dd, J₁=4.4 Hz, J₂=8.0Hz, 1H), 7.33 (dd, J₁=1.6 Hz, J₂=8.8 Hz, 1H), 7.16 (dd, J₁=2.8 Hz,J₂=8.8 Hz, 1H), 7.20 (dd, J₁=2.4 Hz, J₂=8.8 Hz, 1H), 7.04 (dd, J₁=2.8Hz, J₂=8.8 Hz, 1H), 5.80 (d, J=8.8 Hz, 1H), 4.63-4.70 (m, 1H), 4.06-4.13(q, J=7.2 Hz, 2H), 1.46 (t, J=7.2 Hz, 3H), 1.40 (s, 9H), 1.31 (d, J=6.8Hz, 3H) ppm.

Intermediate XVII. To a solution containing 77.00 g (187.59 mmol) of thecompound prepared above in 2.1 L of DCM was added 290 mL trifluoroaceticacid. The reaction was allowed to stir for 3.5 h at room temperaturethen concentrated in vacuo. The concentrate was dissolved in 1.4 L DCMand washed with 1.0 N hydrochloric acid (3×500 ml). The combined aqueouswashes were made alkaline by addition of concentrated ammonium hydroxideuntil pH=10. The resulting cloudy solution was extracted with DCM (2×700ml) and the combined organic extracts dried over magnesium sulfate,filtered, and concentrated in vacuo to afford 58.40 g of product as atan solid. ¹H NMR (DMSO-d₆) δ8.94 (dd, J₁=2.0 Hz, J₂=4.8 Hz, 1H), 8.44(dd, J₃=2.0 Hz, J₂=8.0 Hz, 1H), 7.49 (dd, J₁=4.8 Hz, J₂=8.0 Hz, 1H),7.34-7.39 (m, 2H), 7.04-7.10 (m, 2H), 4.08 (q, J=6.8 Hz, 2H), 3.52 (q,J=6.4 Hz, 1H), 1.94 (br s, 2H), 1.34 (t, J=6.8 Hz, 3H), 1.15 (d, J=6.4Hz, 3H) ppm.

Intermediate XVIII. To a solution containing 57.70 g (185.92 mmol) ofintermediate XVII prepared above dissolved in 1.7 L of dichloroethanewas added 18.5 ml (196.04 mmol) pyridine carboxaldehyde followed by55.20 g (260.45 mmol) of sodium triacetoxy borohydride. The reaction wasallowed to stir at room temperature overnight. The reaction was dilutedwith 1 L of DCM and washed with 1.0 M ammonium hydroxide (2×500 ml). Theorganic phase was dried over magnesium sulfate, filtered, andconcentrated in vacuo to afford 79.20 g of product as a pale yellowsolid. ¹H NMR (DMSO-d₆) δ8.96-8.98 (m, 1H), 8.42-8.48 (m, 1H), 8.45 (brs, 1H), 8.37 (d, J=4.8 Hz, 1H), 7.64 (d, J=8.0 Hz, 1H), 7.52 (dd, J₁=4.8Hz, J₂=8.0 Hz, 1H), 7.33 (dd, J₁=2.4 Hz, J₂=8.4 Hz, 1H), 7.24 (dd,J₃=4.8 Hz, J₂=8.0 Hz, 1H), 7.14 (dd, J₁=2.4 Hz, J₂ =8.4 Hz, 1H), 6.99(dd, J₁=2.8 Hz, J₂=8.4 Hz, 1H), 6.83 (dd, J₃=2.8 Hz, J₂=8.8 Hz, 1H),3.97-4.10 (m, 1H), 3.87 (s, 1H), 3.72 (d, J=14.0 Hz, 1H), 3.52 (d,J=13.6 Hz, 1H), 3.28 (q, J=6.4 Hz, 1H), 1.31 (t, J=7.2 Hz, 3H), 1.17 (d,J=6.4 Hz, 3H) ppm.

Compound 3.16a. To a solution containing 54.00 g (245.29 mmol) of4-(trifluoromethoxy)phenylacetic acid in 1.1 L of DMF was added 61.30 g(319.77 mmol) of EDCI, 43.20 g (319.69 mmol) HOBT and 42.00 ml (382.01mmol) of N-methyl morpholine. After stirring for 30 min., 74.60 g(185.82 mmol) of intermediate XVIII was added. The reaction was allowedto stir at room temperature overnight. The reaction was diluted with 3 LDCM and washed with water (2×3 L), saturated sodium bicarbonate solution(2×2 L), and brine (1×2 L). The organic extract was dried over magnesiumsulfate, filtered, and concentrated in vacuo to afford 121.7 g of ayellow solid. The solids were triturated with 700 ml of methyltert-butyl ether, collected by filtration, rinsed, and dried to afford88.46 g of product as an off-white solid.

The product was recrystallized from 10% ethyl acetate in heptane toafford a colorless, microcrystalline (small needles) solid, m.p. 161.2°C. ¹H NMR (T=120° C.; DMSO-d₆) δ9.01 (dd, J₁=1.6 Hz, J₂=4.4 Hz, 1H),8.46 (dd, J₁=2.0 Hz, J₂=7.6 Hz, 1H), 8.35 (m, 2H), 7.57 (dd, J₁=4.8 Hz,J₂=8.4 Hz, 1H), 7.55 (d, J=8.0 Hz, 1H), 7.43 (d, J=8.0 Hz, 1H),7.06-7.22 (m, 7H), 5.28 (q, J=6.0 Hz, 1H), 4.76 (br s, 2H), 4.13 (q,J=6.8 Hz, 2H), 3.48 (br s, 0.5-1H [H₂O]), 2.91 (br s, 2H), 1.42 (d,J=6.8 Hz, 3H), 1.36 (t, J=6.8 Hz, 3H), ppm. HPLC>99%, chiral HPLC>96%ee). MS (ESI, positive mode): 626 (MH⁺).

Synthesis of Compound 3.16b

Compound 3.16b was prepared following the synthetic procedure forcompound 3.16a, wherein 3-trifluoromethyl-4-fluorophenylacetic acid wasused instead of 4-(trifluoromethoxy)phenylacetic acid.

Synthesis of Compound 3.17

Compound 3.17 was prepared following the synthesis of compound 3.01described above. Example 3.02 was followed for the synthetic sequence,wherein 2-aminonicotinic acid was used in step a instead of2-aminobenzoic acid. Characterization data for compound 3.17 follows:colorless, viscous oil. ¹H NMR similar to spectrum for compound 3.01: amixture of cis/trans amide rotamers in ca. 2:1 (CDCl₃; T=25° C.)characteristic resonance peaks at δ_(minor)5.16 (q, 1.0H, J=6.8 Hz) andδ_(major) 5.40 (q, 2.0H, J=7.2 Hz) ppm. MS (ESI⁺) 588.2 [MH]⁺

Synthesis of Compound 3.17a

Compound 3.17a was prepared following the synthetic procedure forcompound 3.17, wherein 4-trifluoromethylbenzenamine was used instead ofp-phenetidine.

Synthesis of Compound 3.18

Compound 3.18 was prepared following the synthesis of compound 3.01described above. Example 3.02 was followed for the synthetic sequence,wherein 2-aminonicotinic acid was used in step a instead of2-aminobenzoic acid, and 2-bromoethyl ethyl ether was used in step cinstead of 3-picolyl chloride. Characterization data for compound 3.18follows: colorless, viscous oil. ¹H NMR similar to spectrum for compound3.01: a mixture of cis/trans amide rotamers in ca. 3:2 (CDCl₃; T=25° C.)characteristic resonance peaks at δ_(minor)5.00 (q, 1.0H, J=6.4 Hz) andδ_(major)5.00 (q, 1.5H, J=6.8 Hz) ppm. MS (ESI⁺) 569.3 [MH]⁺

Synthesis of Compound 3.19

Compound 3.19 was prepared following the synthesis of compound 3.01described above. Example 3.02 was followed for the synthetic sequence,wherein 2-aminonicotinic acid and 4-cyanoanilne were used in step ainstead of 2-aminobenzoic acid and 4-ethoxyaniline, andimidazole-2-carboxaldehyde was used in step c via reductive aminationinstead of 3-picolyl chloride via amine alkylation. Characterizationdata for compound 3.19 follows: colorless, viscous oil. ¹H NMR singleamide rotamer (CDCl₃; T=25° C.) δ1.45 (d, 3H, J=7.6 Hz), 3.69 (d, 1H,J=15.2 Hz), 3.79 (d, 1H, J=15.2 Hz), 4.74 (q, 1H, J=7.2 Hz), 4.76 (d,1H, J=19.6 Hz), 5.39 (d, 1H, J=19.6 Hz), 7.02 (t, 1H, J=1.6 Hz), 7.07(d, 1H, J=2.0 Hz), 7.14 (d, 2H, J=8.0 Hz), 7.40 (d, 2H, J=8.0 Hz), 7.47(dd, 1H, J₁=2.0 Hz, J₂=8.4 Hz), 7.60 (dd, 1H, J₁=4.8 Hz, J₂=8.0 Hz),7.95 (dd, 1H, J₁=2.0 Hz, J₂=6.8 Hz), 8.00 (dd, 1H, J₁=2.0 Hz, J₂=8.4Hz), 8.11 (dd, 1H, J₁=2.0 Hz, J₂=8.4 Hz), 8.66 (dd, 1H, J₁=1.6 Hz,J₂=7.6 Hz), 9.04 (dd, 1H, J₁=1.6 Hz, J₂=4.4 Hz) ppm. MS (ESI⁺) 569.3[MH]⁺

Synthesis of Compound 3.20

Compound 3.20 was prepared following the synthesis of compound 3.01described above. Example 3.02 was followed for the synthetic sequence,wherein 2-aminonicotinic acid and 4-cyanoaniline were used in step ainstead of 2-aminobenzoic acid and 4-ethoxyaniline, and3-pyridinecarboxaldehyde was used in step c via reductive aminationinstead of 3-picolyl chloride via amine alkylation. Characterizationdata for compound 3.20 follows: colorless, viscous oil. ¹H NMR singleamide rotamer (CDCl₃; T=25° C.) δ1.33 (d, 3H, J=7.2 Hz), 3.66 (d, 1H,J=15.6 Hz), 3.79 (d, 1H, J=15.6 Hz), 5.16 (d, 1H, J=18.0 Hz), 5.19 (q,1H, J=7.2 Hz), 5.24 (d, 1H, J=18.0 Hz), 7.23-7.32 (m, 3H), 7.45 (dd, 1H,J₁=1.6 Hz, J₂=8.0 Hz), 7.49-7.55 (m, 4H), 7.89 (dd, 1H, J₁=1.6 Hz,J₂=8.4 Hz), 7.95 (dd, 1H, J₁=1.2 Hz, J₂=8.0 Hz), 8.02 (dd, 1H, J₁=2.0Hz, H₂=8.4 Hz), 8.52-8.58 (m, 2H), 8.62 (dd, 1H, J₁=2.0 Hz, J₂=8.0 Hz),9.07 (dd, 1H, J₁=2.0 Hz, J₂=4.8 Hz) ppm. MS (ESI⁺) 569.2 [MH]⁺

Synthesis of Compound 3.21

Compound 3.21 was prepared following the synthesis of compound 3.01described above. Example 3.02 was followed for the synthetic sequence,wherein 2-aminonicotinic acid and 4-cyanoaniline were used in step ainstead of 2-aminobenzoic acid and 4-ethoxyaniline, and3-methyl-4-carboxaldehyde-(3H)imidazole was used in step c via reductiveamination instead of 3-picolyl chloride via amine alkylation.Characterization data for compound 3.21 follows: colorless, viscous oil.¹H NMR single amide rotamer (CDCl₃; T=25° C.) δ1.41 (d, 3H, J=7.2 Hz),3.66 (s, 3H), 3.75 (d, 1H, J=16.0 Hz), 3.84 (d, 1H, J=16.0 Hz), 4.98 (s,2H), 5.17 (q, 1H, J=7.2 Hz), 6.86 (s, 1H), 7.30 (s, 1H), 7.40-7.50 (m,3H), 7.50-7.58 (m, 3H), 7.80-8.05 (m, 3H), 8.6 (dd, 1H, J₁=2.0 Hz,J₂=8.0 Hz), 9.06 (dd, 1H, J₁=2.0 Hz, J₂=4.4 Hz) ppm. MS (ESI⁺) 572.2[MH]⁺

Synthesis of Compound 3.22

Compound 3.22 was prepared following the synthetic procedure forcompound 3.02, described above. ¹H NMR (d₆-DMSO, T=140° C.) δ8.13 (d,J=8.1 Hz, 1H), 7.85 (t, J=8.0 Hz, 1H), 7.70 (d, J=8.1 Hz, 1H), 7.65-7.45(m, 5H), 7.40-7.20 (m, 4H), 5.30 (bs, 1H), 4.40 (bs, 2H), 3.40 (bs, 2H),1.99 (s, 3H), 1.34 (d, J=6.6 Hz, 3H). m.p. 220-221° C. MS (ESI⁺) 526.2[MH]⁺.

Synthesis of Compound 3.23

A mixture of compound 3.22 (11 mg, 0.021 mmol) and methoxylaminehydrochloride (0.08 mL, 25-30% aqueous solution) in methanol (4 mL) andpyridine (0.1 mL) was stirred at room temperature for three days. Thesolvents were evaporated, and the residue was purified by column (30%EtOAc in Hexane) to give 12 mg of 3.23 as white solid. MS (ESI⁺) 555.2[MH]⁺.

Synthesis of Compound 3.24

Compound 3.24 was prepared following the synthetic procedure forcompound 3.23, described above. MS (ESI⁺) 541.3 [MH]⁺.

Synthesis of Compound 3.25

Compound 3.25 was prepared following the synthetic procedure forcompound 3.02, described above. ¹H NMR (d₆-DMSO, T=140° C.) δ8.14 (d,J=8.1 Hz, 1H), 7.84 (m, 2H), 7.74 (m, 3H), 7.55 (t, J=8.2 Hz, 1H), 7.30(m, 2H), 7.00 (m, 2H), 5.20 (q, J=6.9 Hz, 1H), 4.05 (dd, J=6.9, 7.0 Hz,2H), 3.80-3.25 (m, 8H), 1.4 (d, J=6.9 Hz, 3H), 1.31 (t, J=7.0 Hz, 3H),0.95 (t, J=7.0 Hz, 3H). m.p. 57-60° C. MS (ESI⁺) 636.2 [MH]⁺. Anal.Calcd. for C₃₂H₃₁F₆N₃O₄: C, 60.47; H, 4.92; N, 6.61. Found C, 60.36; H,4.99; N, 6.51.

Synthesis of Compound 3.26

Compound 3.26 was prepared following the synthetic procedure forcompound 3.02, described above. ¹H NMR (d₆-DMSO, T=140° C.) δ8.13 (d,J=8.0 Hz, 1H), 7.84 (t, J=8.1 Hz, 1H), 7.70 (d, J=8.0 Hz, 1H), 7.54 (t,J=8.2 Hz, 1H), 7.42 (m, 2H), 7.28-7.19 (m, 3H), 7.15 (m, 2H), 5.15 (q,J=6.8 Hz, 2H), 4.08 (q, J=7.0 Hz, 2H), 3.65-3.20 (m, 8H), 1.46 (d, J=6.8Hz, 3H), 1.34 (t, J=7.0 Hz, 3H), 0.96 (t, J=7.0 Hz, 3H). m.p. 137-138°C. MS (ESI⁺) 586.2 [MH]⁺. Anal. Calcd. for C₃₁H₃₁F₄N₃O₄; C, 63.58; H,5.34; N, 7.18. Found: C, 63.47; H, 5.45; N, 7.40.

Synthesis of Compound 3.27

Compound 3.27 was prepared following the synthetic procedure forcompound 3.02, described above. ¹H NMR (d₆-DMSO, T=140° C.) δ8.13 (d,J=8.0 Hz, 1H), 7.84 (t, J=8.1 Hz, 1H), 7.70 (d, J=8.0 Hz, 1H), 7.57 (m,2H), 7.40-7.20 (m, 2H), 7.13 (m, 1H), 7.05 (m, 2H), 6.84 (d, J=8.8 Hz,1H), 5.17 (q, J=6.9 Hz, 2H), 4.09 (q, J=7.0 Hz, 2H), 3.65-3.20 (m, 8H),1.46 (d, J=6.8 Hz, 3H), 1.35 (t, J=7.0 Hz, 3H), 0.96 (t, J=7.0 Hz, 3H).m.p. 146-148° C. MS (ESI⁺) 586.2 [MH]⁺. Anal. Calcd. for C₃₁H₃₁F₄N₃O₄:C, 63.58; H, 5.34; N, 7.18. Found: C, 63.76; H, 5.43; N, 7.19.

Synthesis of Compound 3.28

Compound 3.28 was prepared following the synthetic procedure forcompound 3.02, described above. ¹H NMR (d₆-DMSO, T=140° C.) δ8.33 (m,2H), 8.09 (d, J=8.1 Hz, 1H), 7.84 (t, J=8.0 Hz, 1H), 7.67 (d, J=8.0 Hz,1H), 7.54 (t, J=8.1 Hz, 1H), 7.48 (m, 1H), 7.36 (m, 1H), 7.18-7.04 (m,4H), 7.01 (d, J=8.7 Hz, 2H), 6.92 (d, J=8.7 Hz, 2H), 5.25 (q, J=6.8 Hz,2H), 4.70 (s, 2H), 4.60 (q, J=8.9 Hz, 2H), 4.14 (q, J=7.0 Hz, 2H),3.35-3.00 (bm, 2H), 1.37 (m, 6H). m.p. 103-104° C. MS (ESI⁾ 617.3 [MH]⁺.Anal. Calcd. for 2H), 4.70 (s, 2H), 4.60 (q, J=8.9 Hz, 2H), 4.14 (q,J=7.0 Hz, 2H), 3.35-3.00 (bm, 2H), 1.37 (m, 6H). m.p. 103-104° C. MS(ESI⁾ 617.3 [MH]⁺. Anal. Calcd. for C₃₄H₃₁F₃N₄O₄.1/2H₂O: C, 65.27; H,5.16; N, 8.96. Found: C, 65.01; H, 5.12; N, 8.96.

Synthesis of Compound 3.29

Compound 3.29 was prepared in a manner similar to that used for compound3.02. Light yellow solid, ¹H NMR (DMSO, T=140° C.) 1.35 (t, 3H, J=6.8Hz), 1.50(d, 3H, J=6.8 Hz), 3.58(m, 2H), 4.10(q, 2H, J=6.8 Hz), 4.50 (m,2 H), 5.23 (q, 1H, J=6.8 Hz), 7.11 (m, 2H), 7.29-7.62 (m, 6H), 7.78 (m,2H), 7.88 (t, 1H, J=8 Hz), 8.15 (dd, 1H, J=1.2 Hz, J₂=8.0 Hz). At roomtemperature, mixture of cis/trans amide rotamers (2/1), determined by ¹HNMR (CDCl₃) 5.02 (q, 1H, J=6.8 Hz), 5.51 (q, 1H, J=6.8 Hz). MS(ESI⁺)535.2 (MH⁺).

Synthesis of Compound 3.30

Compound 3.30 was prepared in a manner similar to that used for compound3.02. Light yellow solid, m.p. 153° C. ¹H NMR (DMSO, T=140° C.) 0.97 (t,3H, J=6.8 Hz), 1.37 (t, 3H, J=6.8 Hz), 1.44 (d, 3H, J=6.8 Hz), 3.31-3.59(m, 8H), 4.10 (q, 2H, J=6.8 Hz), 5.17 (q, 1H, J=6.8 Hz), 7.05-7.33 (m,8H), 7.55 (t, 1H, J=8 Hz), 7.71 (d, 1H, J=8 Hz), 7.85 (t, 1H, J=8 Hz),8.15 (dd, 1H, J₁=1.2 Hz, J₂=8.0 Hz). At room temperature, mixture ofcis/trans amide rotamers (1/1), determined by ¹H NMR (CDCl₃) 4.92 (q,1H, J=6.9 Hz), 5.35 (q, 1H, J=6.9 Hz). MS(ESI⁺) 584.3 (MH⁺). Anal.(C₃₁H₃₂F₃N₃O₅) cal. C, 63.80; H, 5.53; N, 7.20. Found C, 63.92; H, 5.61;N, 7.20.

Synthesis of Compound 3.31

Compound 3.31 was prepared in a manner similar to that used for compound3.02. Colorless oil, ¹H NMR (CD₃OD) 1.18 (t, 3H, J=7.0 Hz), 1.37 (t, 3H,J=7.0 Hz), 1.41 (d, 3H, J=6.6 Hz), 3.30 (s, 2H), 3.36 (m, 1H), 3.52 (q,2H, J=7.0 Hz), 3.62 (m, 2H), 3.91 (m, 1H), 4.02 (q, 2H, J=7.0 Hz), 4.75(q, 1H, J=6.6 Hz), 6.85 (m, 1H), 6.90 (d, 2H, J=9 Hz), 7.06 (m, 1H),7.13 (m, 1H), 7.34 (d, 2H, J=4.8 Hz), 7.53 (d, 2H, J=9 Hz), 7.70 (m,1H), 8.28 (d, 2H, J=4.8 Hz) MS(ESI⁺) 501.2 (MH⁺).

Synthesis of Compound 3.32

Compound 3.32 was prepared in a manner similar to that used for compound3.02. Light yellow solid, m.p. 146.3° C. ¹H NMR (DMSO, T=140° C.) 0.97(t, 3H, J=6.8 Hz), 1.36 (t, 3H, J=6.8 Hz), 1.46 (d, 3H, J=6.4 Hz),3.32-3.59 (m, 8H) 4.09 (q, 2H, J=6.8 Hz), 5.17 (q, 1H, J=6.4 Hz),6.95-7.11 (m, 4H), 7.26 (m, 2H), 7.54 (t, 1H, J=8 Hz), 7.71 (d, 1H,J₂=7.6 Hz), 7.85 (dt, 1H, J₁=1.6 Hz, J₂=8.2 Hz), 8.15 (dd, 1H, J=1.2 Hz,J₂=7.6 Hz). At room temperature, mixture of cis/trans amide rotamers(1/1), determined by ¹H NMR (CDCl₃) 4.92 (q, 1H, J=6.8 Hz), 5.38 (q, 1H,J=6.8 Hz). MS(ESI⁺) 554.3 (MH⁺). Anal. (C₃₀H₃₀F₃N₃O₄) cal. C, 65.09; H,5.46; N, 7.59. Found C, 64.93; H, 5.55; N, 7.62.

Synthesis of Compound 3.33

Compound 3.33 was prepared in a manner similar to that used for compound3.02. Light yellow solid, m.p. 77.7° C. ¹H NMR (DMSO, T=140° C.) 1.38(m, 6H), 3.05 (br, 1H), 3.42 (m, 1H), 4.12 (q, 2H, J=7.2 Hz), 4.71 (m,2H), 5.26 (q, 1H, J=6.4 Hz), 7.09-7.51 (m, 9H), 7.39 (m, 1H), 7.51-7.56(m, 2H), 7.67 (d, 1H, J₂=8.4 Hz), 7.85 (t, 1H, J=8 Hz), 8.09 (d, 1H,J=8.0 Hz), 8.34 (m, 1H). At room temperature, mixture of cis/trans amiderotamers (2/1), determined by ¹H NMR (CDCl₃) 5.09 (m, 1H), 5.40 (m, 1H).MS(ESI⁺) 604.2 (MH⁺). Anal. (C₃₃H₂₉ F₃N₄O₄) cal. C, 65.77; H, 4.85; N,9.30. Found C, 65.32; H, 4.87; N, 9.12.

Synthesis of Compound 3.34

Compound 3.34 was prepared in a manner similar to that used for compound3.02. Light yellow solid, m.p. 75.5° C. mixture of cis/trans amiderotamers (2/1), determined by ¹H NMR (CD₃OD) 5.20 (m, 1H), 5.45 (m, 1H).MS(ESI⁺) 556.3 (MH⁺). Anal. (C₃₂H₂₈F₂N₄O₃) cal. C, 69.30; H, 5.09; N,10.10. Found C, 68.83; H, 5.15; N, 9.99.

Synthesis of Compound 3.35

Compound 3.35 was prepared in a manner similar to that used for compound3.02 White solid. ¹H NMR (DMSO, T=140° C.) 1.36(t, 3H, J=6.8 Hz), 1.42(d, 3H, J=6.4 Hz), 3.05 (m, 1H), 3.53(m, 1H), 4.11 (m, 2H), 4.73 (m,2H), 5.27 (q, 1H), J=6.4 Hz), 7.11 (m, 4H), 7.33-7.51 (m, 8H), 7.68 (d,1H, J=8 Hz), 7.83 (t, 1H, J=7.2 Hz), 8.10 (m, 1H), 8.34 (m, 1H). At roomtemperature, mixture of cis/trans amide rotamers (7/1), determined by ¹HNMR (CDCl₃) 5.11 (q, 1H, J=6.4 Hz), 5.42 (m, 1H). MS(ESI⁺) 587.3 (MH⁺).Anal. (C₃₃H₂₉F₃N₄O₃) cal. C, 67.57; H, 4.98; N, 9.55. Found C, 67.15; H,5.12; N, 9.81.

Synthesis of Compound 3.36

Compound 3.36 was prepared in a manner similar to that used for compound3.02. Light yellow solid, ¹H NMR (DMSO, T=140° C.) 1.39 (m, 6H), 3.05(br, 1H), 3.45 (m, 1H), 4.13 (q, 2H, J=6.8 Hz), 4.71 (m, 2H), 5.26 (q,1H, J=6.4 Hz), 7.00-7.16 (m, 8H), 7.35 (m, 2H), 7.37-7.60 (m, 2H), 7.68(d, 1H, J₂=8.4 Hz), 7.84 (t, 1H, J=8 Hz), 8.09 (d, 1H, J=8.0 Hz), 8.34(m, 1H). At room temperature mixture of cis/trans amide rotamers (1/2),determined by ¹H NMR (CDCl₃) 1.25 (d, 1H, J=6.4 Hz), 1.32 (d, 1H, J=6.4Hz). MS(ESI⁺) 603.2 (MH⁺). Anal. (C₃₃H₂₉F₃N₄O₄) cal. C, 65.77; H, 4.85;N, 9.30. Found C, 65.48; H, 4.98; N, 9.39.

Synthesis of Compound 3.37

Compound 3.37 was prepared following the synthesis procedure forcompound 3.02. White solid. ¹H NMR (DMSO, T=120° C.) 1.32 (t, 3H, J=7.07Hz), 1.49-1.55 (m, 6H), 1.70 (m, 1H), 2.26 (m, 1H), 2.58 (m, 1H),2.78-2.88 (m, 4H), 3.12-3.15 (m, 1H), 3.20 (m, 1H), 3.40 (m, 1H), 3.49(m, 1H), 3.50-3.85 (m, 2H), 4.07 (q, 2H, J=7.07 Hz), 5.16 (q, 1H, J=6.67Hz), 7.02 (m, 2H), 7.24 (m, 1H), 7.29-7.44 (m, 4H), 7.56 (t, 1H, J=7.33Hz), 7.72 (d, 1H, J=8 Hz), 7.86 (t, 1H, J=7.6 Hz), 8.14 (d, 1H, J=7.60Hz). MS(ESI⁺) 641.2 (MH⁺).

Synthesis of Compound 3.38

Compound 3.38 was prepared following the synthesis of compound 3.02.Pale yellow solid. ¹H NMR (DMSO, T=120° C.) 1.33 (t, 3H, J=6.67 Hz),1.51 (d, 3H, J=6.67 Hz), 2.95 (m, 1H), 3.19(m, 1H), 3.69 (m, 1H), 3.83(m, 1H), 4.09 (q, 2H, J=6.67 Hz), 5.11-5.23 (m, 3H), 7.06 (m, 2H),7.27-7.45 (m, 7H), 7.56 (t, 1H, J=7.33 Hz), 7.70 (d, 1H, J=8 Hz), 7.85(m, 2H), 8.13 (d, 1H, J=7.60 Hz), 8.48 (s, 1H). MS(ESI⁺) 619.30 (MH⁺).

Synthesis of Compound 3.39.HCl

Compound 3.39 was prepared following the synthesis of compound 3.02.White solid. ¹H NMR (DMSO, T=120° C.) 1.36 (m, 6H), 2.88 (s, 6H), 3.61(d, 1H, J=14.67 Hz), 4.12 (q, 2H, J=6.93 Hz), 4.78 (m, 3H), 5.26 (m,1H), 6.93 (m, 1H), 7.10 (m, 2H), 7.28-7.43 (m, 5H), 7.51-7.59 (m, 3H),7.83 (t, 1H, J=7.33 Hz), 7.97 (m, 1H), 8.08 (d, 1H, J=7.73 Hz). MS(ESI⁺)648.2 (MH⁺).

N-(4-Ethoxy-phenyl)-benzene-1,2-diamine (XIX). In the presence of K₂CO₃(13.0 g, 94.2 mmol), the mixture of 1-fluoro-2-nitrobenzene (8.46 g, 60mmol) and phenylamine (8.22 g, 60 mmol) in DMF (40 ml) was heated to125° C. for 16 h. and then poured into water, the aqueous layer wasextracted with EtOAc three times, the combined organic layer was thenwashed by water, brine and dried over Na₂SO4. The solvent was evaporatedand the crude product was recrystallized from EtOH to afford a brownsolid (4-ethoxy-phenyl)-(2-nitro-pheny)1-amine (10 g).

In the presence of 10% Pd-C (2.1 g, 2 mmol), under hydrogen atmosphere,a solution of (4-ethoxy-phenyl)-(2-nitro-pheny)1-amine (5.16 g, 20 mmol)in MeOH/Et₂O (30 ml/30 ml) was stirred overnight. The solid wasfiltered, the filtrate was evaporated to give a orange solidN-(4-ethoxy-phenyl)-benzene-1,2-diamine (3.6 g) (XIX). ¹H NMR (CDCl₃)1.39 (t, 3H, J=6.93 Hz), 3.98 (q, 2H, J=6.93 Hz), 6.78 (m, 6H), 6.94 (m,1H), 7.03 (m, 1H). MS(ESI⁺) 229.2 (MH⁺).

{1-[2-(4-Ethoxy-phenylamino)-phenylcarbamoyl]-ethyl}-carbamic acidtert-butyl ester (XX). To a solution containing (R)-Boc-Ala-OH (4.92 g,26 mmol) and above diamine (5.4 g, 23.68 mmol) in 50 ml of DMF, wasadded EDCI (9.08 g, 47.3 mmol), HOBt (3.62 g, 23.68 mmol) and NMM (3.59g, 35.52 mmol). The mixture was stirred at room temperature overnight.The reaction was diluted with dichloromethane and washed with water(three times), brine and dried over Na₂SO₄. Removal of solvent affordeda oil which was subjected to flash column. A yellow solid (7.55 g) wasobtained. ¹H NMR (CDCl₃) 1.37 (m, 15H), 3.98 (q, 2H, J=5.2 Hz), 4.21 (m,1H), 4.92 (m, 1H), 5.65 (br, 1H), 6.81 (m, 4H), 6.97 (m, 1H), 7.08 (m,2H), 7.68 (d, 1H, J=8 Hz), 8.16 (s, 1H). MS(ESI⁺) 400.2 (MH⁺).

{1-[1-(4-Ethoxy-phenyl)-1H-benzoimidazol-2-yl]-ethyl}-carbamic acidtert-butyl ester (XXI). A solution of above solid (6 g, 15.03 mmol) inHOAc (60 ml) was heated to 80° C. for 4 h. The solvent was evaporated,the residue was dissolved in EtOAc and washed by sat. NaHCO₃, water,brine and dried over Na₂SO4. The solvent was removed and the cruderesidue was subjected by flash column to afford a white solid (4.1 g).¹H NMR (CDCl₃) 1.36 (s, 9H), 1.43 (m, 6H), 4.10 (q, 2H, J=6.93 Hz), 4.84(m, 1H), 7.03 (m, 1H), 7.06 (m, 2H), 7.21 (m, 2H), 7.38 (m, 2H), 7.66(d, 1H, J=7.73 Hz). MS(ESI⁺) 382.3 (MH⁺).

Synthesis of Compound 3.40.HCl

Starting from{1-[1-(4-ethoxy-phenyl)-1H-benzoimidazol-2-yl]-ethyl}-carbamic acidtert-butyl ester, compound 3.40 was prepared following the synthesis ofcompound 3.02. Yellow solid. ¹H NMR (DMSO, T=120° C.) 1.35 (t, 3H,J=6.93 Hz), 1.66 (m, 3H), 3.29 (br, 1H), 3.93 (d, 1H, J=16 Hz), 4.11 (q,2H, J=6.93 Hz), 4.65 (m, 1H), 4.75 (m, 1H), 5.70 (m, 1H), 7.03 (d, 1H,J=7.73 Hz), 7.14 (d, 2H, J=8.27 Hz), 7.26-7.44 (m, 5H), 7.70-7.84 (m,5H), 8.45 (s, 2H). MS(ESI⁺) 627.2 (MH⁺).

Synthesis of Compound 3.41

Compound 3.41 was prepared following the synthesis of compound 3.40.White solid. ¹H NMR (DMSO, T=120° C.) 1.41 (t, 3H, J=6.93 Hz), 1.59 (m,3H), 3.30 (br, 1H), 3.69 (d, 1H, J=16 Hz), 4.15 (q, 2H, J=6.93 Hz),4.49-4.63 (m, 2H), 5.74 (m, 1H), 7.02-7.15 (m, 6H), 7.30-7.41 (m, 3H),7.58 (m, 2H), 8.18-8.32 (m, 3H). MS (ESI⁺) 602.3 (MH⁺).

Synthesis of Compound 3.42.HCl

Compound 3.42 was prepared following the synthesis of compound 3.02.White solid. ¹H NMR (DMSO, T=120° C.) 1.35 (t, 3H, J=6.93 Hz), 1.52 (m,3H), 1.9 (m, 6H), 2.85-3.05 (m, 3H), 3.36-3.57 (m, 4H), 3.95 (m, 3H),4.08 (q, 2H, J=6.93 Hz), 5.17 (m, 1H), 7.02-7.10 (m, 2H), 7.24-7.46 (m,5H), 7.58 (t, 1H, J=7.6 Hz), 7.75 (d, 1H, J=7.73 Hz). 7.88 (t, 1H,J=7.73 Hz), 8.15 (d, 1H, J=8 Hz). MS(ESI⁺) 625.3 (MH⁺).

Synthesis of Compound 3.43

4-{1-[3-(4-Ethoxy-phenyl)-4-oxo-3,4-dihydro-quinazolin-2-yl]-ethylamino}-piperidine-1-carboxylicacid tert-butyl ester (I). 4-Oxo-Piperidine-1-carboxylic acid tert-butylester (0.468 g, 2.35 mmol) was added to a solution of amine (0.6g, 1.96mmol) in dichloroethane (10 ml) at −10° C., followed by Na(OAC)₃BH(0.602 g, 2.84 mmol). The mixture was kept at that temperature for 1.5h, then warmed slowly to room temperature and stirred overnight. Thesolution was diluted with DCM, washed by sat.NaHCO₃, water, brine anddried over Na₂SO₄ The solvent was evaporated and a white solid (1.1 g)was obtained, which was used in the next step. MS(ESI⁺) 493.3 (MH⁺).

4-{{1-[3-(4-Ethoxy-phenyl)-4-oxo-3,4-dihydro-quinazolin-2-yl]-ethyl}-[(4-fluoro-3-trifluoromethyl-phenyl)-acetyl]-amino}-piperidine-1-carboxylicacid ester (II). Pyridine (0.289 g, 3.66 mmol) was added to a mixture ofI (0.6 g, 1.22 mmol) and phenylacetyl chloride (0.44 g, 1.83 mmol) intoluene (15 ml). The solution was heated to 60° C. for 3 h, and thenpoured into 1N HCl. The aqueous layer was extracted with EtOAc threetimes, the combined organic layer was then washed by sat. NaHCO₃, water,brine and dried over Na₂SO4. The solvent was evaporated and the crudeoil was subjected by flash column to afford a yellow solid (540 mg).MS(ESI⁺) 697.3 (MH⁺).

{1-[3-(4-Etboxy-phenyl)-4-oxo-3,4-dihydro-quinazolin-2-yl]-ethyl}-2-(4-fluoro-3-trifluoromethyl-phenyl)-piperidin4-yl-acetamide(3.43). Trifluoroacetic acid (1.77 g, 15.5 mmol) was added to a solutionof II (0.54 g, 0.77 mmol) in dichloromethane. The mixture was stirred atroom temperature for 3 h. The solvent was evaporated, the residue wasdissolved in EtOAc and washed with sat. NaHCO₃, water, brine and driedover Na₂SO4. The solvent was evaporated and the crude oil was subjectedby flash column to afford a white solid (440 mg). ¹H NMR (DMSO, T=120°C.) 1.21 (m, 1H), 1.34 (m, 4H), 1.48 (d, 3H, J=6.8 Hz), 2.16 (m, 3H),2.70-2.97 (m, 4H), 3.28 (d, 1H, J=16 Hz), 3.64 (m, 1H), 4.09 (q, 2H,J=6.8 Hz), 5.04 (m, 1H), 7.07-7.20 (m, 3H), 7.30-7.44 (m, 4H), 7.58 (t,1H, J=7.33 Hz), 7.76 (d, 1H, J=8 Hz), 7.88 (d, 1H, J=7.07 Hz), 8.14 (d,1H, J=7.73 Hz), MS(ESI⁺) 597.3 (MH⁺).

Synthesis of Compound 3.44

Formaldehyde (37% in water) (0.016 g, 0.20 mmol) was added to a solutionof T0913409 (0.06 g, 0.1 mmol) in dichloroethane (5 ml), followed byNa(OAC)₃BH (0.127 g, 0.60 mmol) at room temperature. The mixture wasstirred overnight. The solution was diluted with DCM, washed bysat.NaHCO₃, water, brine and dried over Na₂SO₄. The solvent wasevaporated and the residue was purified by flash column to afford awhite solid (58 mg). ¹H NMR (DMSO, T=120° C.) 1.18 (m, 1H), 1.33 (t, 3H,J=6.8 Hz), 1.48 (d, 3H, J=6.67 Hz), 1.61 (m, 1H), 1.95 (m, 1H), 2.10 (s,3H), 2.38 (m, 2H), 2.64 (m, 1H), 2.76 (m, 1H), 2.88 (s, 2H), 3.26 (d,1H, J=16 Hz), 3.58 (m, 1H), 4.09 (d, 2H, J=6.8 Hz), 5.04 (m, 1H),7.08-7.45 (m, 7H), 7.58 (m, 1H), 7.76 (d, 1H, J=8 Hz), 7.89 (m, 1H),8.14 (d, 1H, J=7.73 Hz). MS(ESI⁺) 611.3 (MH⁺).

Synthesis of Compound 3.45

3-(4-Ethoxyphenyl)-2-ethyl-quinoline. 33% Potassium hydroxide (1.3 ml)was added to a mixture of o-amino aldehyde (0.31 g, 2.6 mmol) and ketone(0.5 g, 2.6 mmol) in 95% EtOH. The solution was heated to reflux for 2 hand then poured into water. The aqueous layer was extracted with EtOActhree times, the combined organic layer was then washed by water, brineand dried over Na₂SO4. The solvent was evaporated and the crude oil wassubjected by flash column to afford a white solid (170 mg). ¹H NMR(CDCl₃) 1.23 (t, 3H, J=7.5 Hz), 1.47 (t, 3H, J=6.93 Hz), 2.98 (q, 2H,J=7.47 Hz), 4.11 (q, 2H, J=6.93 Hz), 6.98 (m, 2H, J=6.8 Hz), 7.31 (m,2H), 7.49 (m, 1H), 7.68 (m, 1H), 7.77 (d, 1H, J=8 Hz), 7.92 (s, 1H),8.08 (d, 1H, J=8 Hz). MS(ESI⁺) 278.3 (MH⁺).

Starting from 3-(4-ethoxy-phenyl)-2-ethyl-quinoline, compound 3.45 wasprepared following the synthesis of compound 1.01 (IV-1.01). yellowsolid. ¹H NMR (DMSO, T=120° C.) 1.35 (t, 3H, J=6.80 Hz), 1.57 (m, 3H),2.94 (br, 1H), 3.63 (d, 1H, J=16 Hz), 4.11 (q, 2H, J=6.80 Hz), 4.63 (m,2H), 5.88 (m, 1H), 7.07 (m, 5H), 7.22-7.33 (m, 3H), 7.58 (m, 2H), 7.76(t, 1H, J=7.6 Hz), 7.90 (d, 1H, J=8 Hz), 7.08-8.12 (m, 3H), 8.24 (m,1H). MS(ESI⁺) 588.3 (MH⁺).

Synthesis of Compound 3.46

(R)-2-((N-3-Picolyl)-N-(4-trifluoromethylphenylacetyl)-1-aminoethyl)-3-(4-trifluoroethyl)-3H-quinazoline-4-one (3.46). Compound 1 wassynthesized using the method described in FIG. 4, except that4-trifluoromethylaniline was used in place of p-phenetidine. ¹H NMR(DMSO-d₆, T=120° C.) 1.40 (m, 3H), 2.89 (m, 1H), 3.58 (m, 1H), 4.78 (m,2H), 5.24 (m, 1H), 7.21 (d, J=4.0 Hz, 1H), 7.26 (m, 2H), 7.55 (m, 5H),7.71 (m, 1H), 7.80 (br s, 1H), 7.90 (m, 3H), 8.12 (d, J=7.8 Hz, 1H),8.38 (t, J=5.1 Hz, 2H) ppm. MS (ESI⁺) m/z 611.3 [M+H]⁺.

Synthesis of Compound 3.47

(R)-2-((N-3-Picolyl)-N-(4-trifluoromethylphenylacetyl)-1-aminoethyl)-3-(4-(1-propynyl))-3H-quinazoline-4-one (3.47). Compound 3.47 wassynthesized using the method described for the synthesis of compound3.07, except that excess amount of propyne gas was used in place oftrimethylsilylacetylene. ¹H NMR (DMSO-d₆, T=120° C.) 1.41 (m, 3H), 2.09(s, 3H), 2.89 (m, 1H), 3.55 (m, 1H), 4.72 (m, 2H), 5.23 (m, 1H), 7.18(m, 1H), 7.28 (m, 2H), 7.55 (m, 8H), 7.71 (m, 1H), 7.87 (m, 1H), 8.11(d,J=7.8 Hz, 1H), 8.37 (m, 2H) ppm. MS (ESI⁺) m/z 581.2 [M+H]⁺.

Synthesis of Compound 3.48

(R)-2-((N-3-Picolyl)-N-(4-trifluoromethylphenylacetyl)-1-aminoethyl)-3-(4-carboethoxymethoxy)-3H-quinazoline-4-one (3.48). Compound 3.48 wassynthesized using the method described in FIG. 4, except that4-(carboethoxymethoxy)aniline was used in place of p-phenetidine. ¹H NMR(DMSO-d₆, T=120° C.) 1.24 (t, J=7.1 Hz, 3H), 1.39 (d, J=5.4 Hz, 3H),3.51 (br s, 1H), 4.21 (q, J=7.1 Hz, 2H), 4.72 (br s, 2H), 4.78 (s, 2H),5.22 (m, 2H), 7.16 (m, 3H), 7.29 (d, J=7.7 Hz, 2H), 7.44 (m, 1H), 7.54(m, 4H), 7.68 (m, 2H), 7.86 (t, J=7.0 Hz, 1H), 8.10 (d, J=7.0 Hz, 1H),8.36 (br s, 2H) ppm. MS (ESI⁺) m/z 645.2 [M+H]⁺.

Synthesis of Compound 3.49

4-(2,2,2-trifluoroethoxy)aniline. To a mixture of 4-nitrophenol (1.39 g,10 mmol, 1.0 equiv), and K₂CO₃ (1.8 g, 13 mmol, 1.3 equiv) in 10 mL ofdry DMF was added 1-iodo-2,2,2-trifluoroethane (2.31 g, 11 mmol, 1.1equiv). The mixture was heated in an oil bath at 100° C. for 24 h. Halfof the initial amount of K₂CO₃ and 1-iodo -2,2,2-trifluoroethane wereadded. The mixture was stirred for another 24 h at 100° C. This wasrepeated once more on the third day. At the end of this 72 h reaction,the mixture was cooled to room temperature and poured into 40 mL ofwater. The mixture was extracted twice with 20 mL of diethyl ether. Thecombined ether extract was washed once with 40 mL of brine, dried overanhydrous Na2SO4, filtered to remove drying agent, and evaporated invacuo to yield 1.6 g of a crude product as light yellow solid. ¹H NMR(DMSO-d₆) 4.98 (q, J=8.8 Hz, 2H), 7.30 (d, J=9.2 Hz, 2H), 8.26 ((d,J=9.2 Hz, 2H) ppm.

To a solution of the crude 4-(2,2,2-trifluoroethoxy)nitrobenzene (1.6 g,7.2 mmol, 1.0 equiv) in 40 mL of dichloromethane was added 0.4 g of a 5%palladium on activated carbon (0.19 mmol, 0.026 equiv). Hydrogen gas wasintroduced using a balloon while the mixture was stirred vigorously for48 h at room temperature. After all starting material had been consumed,the mixture was filtered through a pad of Celite to remove the palladiumcatalyst. The filtrate was evaporated in vacuo to give a crude productas a brown liquid. This crude product was purified by distillation atreduced pressure to give 1.3 g of pure 4-(2,2,2-trifluoroethoxy)anilineas a colorless liquid, which solidified upon cooling to 0° C. b.p.81-83° C. at 0.5 torr;. ¹H NMR (DMSO-d₆) δ4.53 (q, J=9.1 Hz, 2H), 4.76(br s, 2H), 6.52 (d, J=7.6 Hz, 2H), 6.76 (d, J=7.6 Hz, 2H) ppm.

(R)-2-((N-3-Picolyl)-N-(3,4-dichlorophenylacetyl)-1-aminoethyl)-3-(4-(2,2,2-trifluoroetboxy)phenyl)-3H-quinazoline-4-onetrifluoroacetate (3.49.CF₃COOH). The trifluoroacetic acid salt ofcompound 3.49 was synthesized using the method described in FIG. 4,except that 4-(2,2,2-trifluoroethoxy)aniline was used in place ofp-phenetidine, and that 3,4-dichlorophenylacetic acid was used in placeof 4-trifluoromethylphenylacetic acid. ¹H NMR (DMSO-d₆, T=120° C.) δ1.42(br s, 3H), 3.50 (m, 1H), 4.75 (m, 5H), 5.22 (m, 1H), 7.03 (d, J=7.9 Hz,1H), 7.27 (m, 5H), 7.43 (d, J=8.2 Hz, 1H), 7.50 (m, 1H), 7.56 (t, J=7.4Hz, 1H), 7.67 (t, J=8.4 Hz, 2H) 7.87 (t, 1H), 8.11 (d, J=7.6 Hz, 1H),8.42 (br s, 2H) ppm. MS (ESI⁺) m/z 641.2 [M+H]⁺.

Synthesis of Compound 3.50

(R)-2-((N-3-Picolyl)-N-(4-trifluoromethoxyphenylacetyl)-1-aminoethyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-3H-8-azaquinazoline-4-one(5). Compound 3.50 was synthesized using the method described in FIG.13, except that 4-(2,2,2-trifluoroethoxy)aniline was used in place ofp-phenetidine. ¹H NMR (DMSO-d₆, T=120° C.) δ1.42 (m, 3H), 3.51 (m, 1H),4.17 (m, 1H), 4.77 (q, J=8.7 Hz, 2H), 4.89 (m, 2H), 5.25 (m, 1H), 7.18(m, 4H), 7.28 (m, 3H), 7.47 (m, 1H), 7.59 (m, 2H), 7.90 (m, 1H), 8.50(m, 3H), 9.01 (m, 1H) ppm. MS (ESI⁺) m/z 658.2 [M+H]⁺.

Synthesis of Compound 3.51

(R)-2-((N-3-Picolyl)-N-(4-trifluoromethoxyphenylacetyl)-1-aminoethyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-3H-5,6,7,8-tetrahydro-8-azaquinazoline-4-one(3.51). To a solution of compound 3.50 (10 mg, 15 μmol, 1.0 equiv) in1.0 mL of MeOH, was added 10% Pd on activated carbon (2 mg, 1.9 μmol,0.13 equiv). Hydrogen was introduced using a balloon. The mixture wasstirred vigorously for 16 h at room temperature. The mixture was dilutedwith 5 mL of dichloromethane and filtered to removed catalyst. Thefiltrated was evaporated in vacuo to give crude 6, which was purified bysilica gel chromatography to give 7.3 mg 3.51 as a white solid. ¹H NMR(DMSO-d₆, T=120° C.) δ1.26 (d, J=6.5 Hz, 3H), 1.79 (m, 2H), 2.34 (t,J=6.1 Hz, 2H), 2.88 (m, 1H), 3.29 (m, 3H), 4.62 (m, 2H), 4.70 (q, J=8.8Hz, 2H), 5.07 (m, 1H), 6.43 (s, 1H), 7.00 (m, 1H), 7.16 (m, 6H), 7.29(m, 2H), 7.51(d, 1H), 8.41(m, 2H) ppm. MS (ESI⁺) m/z 662.2 [M+H]⁺.

Synthesis of Compound 3.52

Compound 3.52, white solid. ¹H NMR (DMSO, T=120° C.) 8.37-8.41(m, 2H),8.12 (d, 1H, J=7.2 Hz), 7.99 (m, 2H), 7.89 (m, 1H), 7.79 (m, 1H), 7.69(d, 1H, J=7.0 Hz), 7.53-7.61 (m, 4H), 7.09-7.23 (m, 3H), 5.23 (m, 1H),4.68-4.81 (m, 2H), 3.65-3.70 (m, 1H), 2.96-3.22 (m, 1H), 1.40 (m, 3H).MS (ESI⁺) 586.2 (MH⁺).

Synthesis of Compound 3.53

Compound 3.53, white solid. ¹H NMR (DMSO, T=120° C.) 8.35 (m, 2H), 8.10(d, 1H, J=6.9 Hz), 7.86 (t, 1H, J=7.0 Hz), 7.70 (m, 1H), 7.41-7.31 (m,3H), 7.29 (m, 2H), 7.16 (m, 8H), 6.94-7.00 (m, 3H), 5.26 (m, 1H), 4.71(m, 2H), 4.36-4.41 (m, 4), 3.44 (m, 1H), 3.05 (m, 1H), 1.39 (m, 3H). MS(ESI⁺) 695.2 (MH⁺).

Synthesis of Compound 3.54

Compound 3.54, white solid. ¹H NMR (DMSO, T=120° C.) 8.10 (d, 1H, J=7.5Hz), 7.86 (t, 1H, J=7.1 Hz), 7.40-7.71(m, 9H), 7.10 (m, 4H), 5.25 (m,1H), 4.77 (m, 2H), 4.12 (q, 2H, J=7.0 Hz), 3.61 (m, 1H), 3.05 (m, 1H),1.43 (m, 3H), 1.36 (t, 3H, J=7.0 Hz). MS (ESI⁺) 629.2 (MH⁺).

Synthesis of Compound 3.55

Compound 3.55, white solid. ¹H NMR (DMSO, T=120° C.) 8.10 (d, 1H, J=8.0Hz), 7.86 (t, 1H, J=7.2 Hz), 7.71 (m, 1H), 7.54-7.60 (m, 2H), 7.37 (m,1H), 7.03-7.11 (m, 5H), 6.76 (m, 1H), 6.70 (m, 2H), 5.28 (m, 1H),4.61-4.63 (m, 2H), 4.10 (q, 2H, J=7.0 Hz), 3.70 (s, 3H), 3.65 (m, 1H),3.59 (s, 3H), 2.95 (m, 1H), 1.43 (m, 3 H), 1.36 (q, 3H, J=7.0). MS(ESI⁺) 664.2 (MH⁺).

Synthesis of Compound 3.56

Compound 3.56, white solid. ¹H NMR (DMSO, T=120° C.) 9.06 (s, 1H), 8.70(d, 1H, J=5.1 Hz), 8.35-8.37 (m, 2H), 7.90 (d, 1H, J=5.1 Hz), 7.55 (m,1H), 7.41 (m, 3H), 7.28-7.33 (m, 1H), 7.07-7.19 (m, 4H), 5.28 (m, 1H),4.74 (m, 2H), 4.10 (q, 2H, J=7.0 Hz), 3.60 (m, 1H), 2.86 (m, 1H), 1.45(m, 3H), 1.35 (t, 3H, J=7.0 Hz). MS (ESI⁺) 606.2 (MH⁺).

Synthesis of Compound 3.57

Compound 3.57, white solid. ¹H NMR (DMSO, T=120° C.) 8.36 (m, 2H), 8.10(s, 1H, J=8.1 Hz), 7.87 (t, 1H, J=7.0 Hz), 7.71 (m, 1H), 7.47-7.61 (m,6H), 7.07-7.19 (m, 4H), 5.23 (m, 1H), 4.71 (m, 2H), 4.50 (s, 2H), 3.58(m, 1H), 3.37 (s, 3H), 2.90 (m, 1H), 1.43 (m, 3H). MS (ESI⁺) 605.3(MH⁺).

Synthesis of Compound 3.58

Compound 3.58, white solid. ¹H NMR (DMSQ, T=120° C.) 8.20 (d, 1H, J=1.0Hz), 8.17 (s, 1H), 8.09 (m, 1H), 7.87 (m, 1H), 7.72 (m, 1H), 7.56 t, 1H,J=7.2 Hz), 7.38-7.40 (m, 3H), 7.29-7.34 (m, 1H), 7.17 (m, 1H), 7.03-7.11(m, 3H), 5.27 (m, 1H), 4.68-4.71 (m, 2H), 4.09 (q, 2H, J=7.0 Hz),3.52-3.58 (m, 1H), 2.90 (m, 1H), 2.07 (s, 3H), 1.44-1.47 (m, 3H), 1.34(t, 3H, J=7.0 Hz). MS (ESI⁺) 619.2 (MH⁺).

Synthesis of Compound 3.59

Compound 3.59, white solid. ¹H NMR (DMSO, T=120° C.) 8.07 (d, 1H, J=8.0Hz), 7.85 (t, 1H, J=6.9 Hz), 7.70 (m, 1H), 7.54 (m, 1H), 7.27-7.38 (m,4H), 7.04-7.09 (m, 7H), 5.28 (m, 1H), 4.68 (m, 2H), 4.09 q, 2H, J=6.9Hz), 3.49-3.58 (m, 3H), 2.90 (m, 1H), 2.46-2.51 (m, 4H), 1.43 (m, 3H),1.35 (t, 3H, J=6.9 Hz), 0.96 (t, 6H, J=7.0 Hz). MS (ESI⁺) 688.5 (MH⁺).

Synthesis of Compound 3.60

Compound 3.60 was prepared as outlined in Scheme 11, below. White solid.¹H NMR (DMSO, T=120° C.) 9.00 (m, 1H), 8.51 (m, 1H), 7.58 (m, 1H), 7.41(m, 1H), 7.18-7.26 (m, 5H), 7.04-7.10 (m, 2H), 5.16 (m, 1H), 4.11 (q,2H, J=7.0 Hz), 3.40-3.50 (m, 7H), 2.90 (m, 1H), 2.30-2.40 (m, 6H), 1.46(m, 3H), 1.36 (t, 3H, J=7.0 Hz). MS (ESI⁺) 626.4 (MH⁺).

Synthesis of Compound 3.61

3.61 was synthesized following the generic synthetic scheme for thesynthesis of triazolinones (FIG. 9) to yield a colorless solid. ¹H NMR(d₆-DMSO; T=120° C.) δ8.57 (d, J=14 Hz, 2H), 7.91 (s, 1H), 7.90 (d,J=8.0 Hz, 2H), 7.58 (s, 1H), 7.57 (d, J=8.4 Hz, 2H), 7.41 (t, J=7.6 Hz,2H), 7.35 (d, J=8.4 Hz, 2H), 7.27 (t, J=7.6 Hz, 1H), 7.24 (d, J=7.6 Hz,2H), 7.09 (d, J=8.8 Hz, 2H), 5.50 (s, 1H), 4.73 (d, J=16.8 Hz, 1H), 4.63(d, J=16.8 Hz, 1H), 4.12 (q, J=6.8 Hz, 2H), 3.67 (d, J=16.0 Hz, 1H),3.31 (br s, 1H), 1.48 (d, J=6.8 Hz, 3H), 1.37 (t, J=6.8 Hz, 3H) ppm.MS(ESI⁺⁾ 602.2 (MH⁺).

Synthesis of Compound 3.62

3.62 was synthesized following the synthetic scheme for8-azaquinazolinones (FIG. 13) to yield a faint yellow solid. ¹H NMR(d₆-DMSO; T=120° C.) 9.02 (dd, J₁=2.0 Hz, J₂=4.4 Hz, 1H), 8.57 (br s,1H), 8.51 (br s, 1H), 8.50 (dd, J₁=1.6 Hz, J₂=8.0 Hz, 1H), 8.02 (m, 2H),7.94 (br s, 1H), 7.83 (br s, 1H), 7.67 (br s, 1H), 7.60 (dd, J₁=4.4 Hz,J₂=7.6 Hz, 1H), 7.50 (br s, 1H), 7.44 (d, J=8.4 Hz, 1H), 7.42 (d, J=6.8Hz, 1H), 7.30 (dd, J₁=J₂=8.8 Hz, 1H), 5.23 (q, J=6.4 Hz, 1H), 4.96 (d,J=18.0 Hz, 1H), 4.86 (d, J=18.0 Hz, 1H), 3.70 (d, J=16.4 Hz, 1H), 3.39(br s, 1H), 1.42 (d, J=6.4 Hz, 3H) ppm. MS(ESI⁺) 587.3 (MH⁺).

Synthesis of Compound 3.63

Compound 3.63 was synthesized following the synthetic scheme for8-azaquinazolinones (FIG. 13) to yield a colorless solid. ¹H NMR(d₆-DMSO; T=120° C.) δ9.01 (dd, J₁=1.6 Hz, J₂=4.4 Hz, 1H), 8.44 (dd,J₁=1.6 Hz, J₂=7.6 Hz, 1H), 7.55 (dd, J₁=4.8 Hz, J₂=8.0 Hz, 1H), 7.48 (d,J=8.0 Hz, 1H), 7.35-7.44 (m, 3H), 7.32 (d, J=8.4 Hz, 1H), 7.27 (d, J=8.8Hz, 1H), 6.93-7.12 (m, 5H), 6.87 (dd, J₁=J₂=4.0 Hz, 1H), 5.25 (q, J=6.4Hz, 1H), 4.82 (d, J=16.4 Hz, 1H), 4.71 (d, J=16.8 Hz, 1H), 4.09 (q,J=7.2 Hz, 2H), 3.64 (d, J=15.6 Hz, 1H), 2.98 (br m, 1H), 1.49 (d, J=6.8,3H), 1.35 (t, J=6.8 Hz, 3H) ppm. MS(ESI⁺) 666.2 (MNa⁺).

Synthesis of Compound 3.64

Compound 3.64 was synthesized following the synthetic scheme for8-azaquinazolinones (FIG. 13) to yield a yellow glassy solid. ¹H NMR(d₆-DMSO; T=120° C.) δ9.02 (dd, J₁=1.6 Hz, J₂=4.4 Hz, 1H), 8.52 (dd,J₁=2.0 Hz, J₂=10.4 Hz, 7.59 (dd, J₁=4.4 Hz, J₂=7.6 Hz, 1H), 7.38-7.52(m, 3H), 7.32 (dd, J₁=J₂=10.4 Hz, 1H), 7.28 (d, J=9.2 Hz, 1H), 6.90-7.04(m, 2H), 5.14 (q, J=6.4 Hz, 1H), 4.09 (q, J=6.8 Hz, 2H), 3.48-3.72 (brm, 3H), 3.02-3.17 (br m, 4H), 2.77-2.98 (br m, 5H), 2.67 (s, 3H), 1.51(d, J=6.4 Hz, 3H), 1.35 (t, J=6.8 Hz, 3H) ppm. MS(ESI⁺) 641.3 (MH⁺).

Synthesis of Compound 3.65

Compound 3.65 was synthesized following the synthetic scheme for thegeneric synthesis of 8-azaquinazolinones (FIG. 13) to yield a colorlesssolid. ¹H NMR (d₆-DMSO; T=120° C.) δ9.03 (d, J=2.8, 1H), 8.50 (dd,J₁=1.6 Hz, J₂=7.6 Hz, 1H), 7.59 (dd, J₁=4.8 Hz, J₂=7.6 Hz, 1H), 7.43 (d,J=8.4 Hz, 1H), 7.38 (d, J=6.4 Hz, 1H), 7.34-7.47 (m, 1H), 7.09 (d, J=8.0Hz, 1H), 6.99 (d, J=7.2 Hz, 1H), 6.66-6.92 (m, 3H), 5.26 (br s, 1H),4.83 (br s, 2H), 4.08 (q, J=7.2 Hz, 2H), 3.56 (br s, 1H), 2.98 (br s,1H), 1.52 (d, J=6.8 Hz, 3H), 1.33 (t, J=6.8 Hz, 3H) ppm. MS(ESI⁺) 611.2(MH⁺), 633.2 (MNa⁺).

Synthesis of Compound 3.66

Compound 3.66 was synthesized following the synthetic scheme for thegeneric synthesis of 8-azaquinazolinones (FIG. 13) to yield a yellowglassy solid. ¹H NMR (d₆-DMSO; T=120° C.) δ9.00 (d, J=2.4 Hz, 1H), 8.52(d, J=7.6 Hz, 1H), 7.90 (dd, J₁=4.4 Hz, J₂=7.6 Hz, 1H), 7.41-7.54 (m,3H), 7.37 (d, J=11.6 Hz, 1H), 7.32 (dd, J₁=J₂=9.2 Hz, 1H), 7.04-7.18 (m,2H), 5.10 (br s, 1H), 4.12 (q, J=7.2 Hz, 2H), 3.90-4.06 (m, 1H),3.62-3.84 (m, 1H), 3.40-3.60 (m, 1H), 2.96-3.14 (m, 1H), 2.74 (s, 3H),1.52 (d, J=6.4 Hz, 3H), 1.36 (s, 9H), 1.34 (t, J=7.2 Hz, 3H) ppm.MS(ESI⁺) 628.4 (MH⁺).

Synthesis of Compound 3.67

Compound 3.67 was synthesized following the synthetic scheme for thegeneric synthesis of 8-azaquinazolinones (FIG. 13) to yield a colorlesssolid. ¹H NMR (d₆-DMSO; T=120° C.) δ9.04 (dd, J₁=1.6 Hz, J₂=4.4 Hz, 1H),8.50 (dd, J₁=2.0 Hz, J₂=7.6 Hz, 1H), 7.58 (dd, J₁=4.4 Hz, J₂=8.0 Hz,1H), 7.40 (d, J=6.8 Hz, 2H), 7.36-7.45 (m, 1H), 7.30 (dd, J₁=J₂=10.4 Hz,1H), 7.27-7.34 (m, 1H), 7.08 (d, J=8.4 Hz, 2H), 7.02 (d, J=6.8 Hz, 1H),5.23 (br s, 1H), 4.50 (d, J=15.6 Hz, 1H), 4.43 (d, J=15.6 Hz, 1H), 4.09(q, J=6.8 Hz, 1H), 3.67 (s, 3H), 2.93 (br s, 2H), 1.49 (d, J=6.4 Hz,3H), 1.35 (t, J=6.8 Hz, 3H) ppm. MS(ESI⁺) 609.3 (MH⁺), 631.2 (MNa⁺).

Synthesis of Compound 3.68

Compound 3.68 was synthesized following the synthetic scheme for thegeneric synthesis of 8-azaquinazolinones (FIG. 13) to yield a yellowglassy solid. ¹H NMR (d₆-DMSO; T=120° C.) δ9.00 (dd, J₁=2.0 Hz, J₂=4.4Hz, 1H), 8.44 (dd, J₁=2.4 Hz, J₂=8.4 Hz, 1H), 7.56 (dd, J₁=4.8 Hz,J₂=8.0 Hz, 1H), 7.33-7.44 (m, 3H), 7.28 (dd, J₁=J₂=10.4 Hz, 1H), 7.06(dd, J₁=J₂=8.8 Hz, 1H), 7.05 (d, J=5.2 Hz, 2H), 6.95 (d, J=8.4 Hz, 2H),6.52 (d, J=7.2 Hz, 2H), 5.27 (q, J=6.4 Hz, 1H), 4.65 (d, J=16.4 Hz, 1H),4.48 (d, J=16.4 Hz, 1H), 4.10 (q, J=6.8 Hz, 2H), 3.58 (d, J=15.2 Hz,1H), 2.90 (br s, 1H), 2.82 (s, 3H), 1.45 (d, J=6.4 Hz, 3H), 1.35 (t,J=6.8 Hz, 3H) ppm. MS(ESI⁺) 670.3 (MNa⁺).

Synthesis of Compound 3.69

Compound 3.69 was synthesized following the synthetic scheme for thegeneric synthesis of 8-azaquinazolinones (FIG. 13) to yield a colorlesssolid. ¹H NMR (d₆-DMSO; T=120° C.) δ9.01 (dd, J₁=2.0 Hz, J₂=4.4 Hz, 1H),8.47 (dd, J₁=2.0 Hz, J₂=8.0 Hz, 1H), 8.36 (s, 2H), 7.58 (dd, J₁=4.4 Hz,J₂=8.0 Hz, 1H), 7.44 (d, J=7.2 Hz, 1H), 7.08-7.22 (m, 7H), 5.26 (q,J=6.8 Hz, 1H), 4.68 (br s, 2H), 4.13 (q, J=7.2 Hz, 2H), 2.89 (br s, 2H),2.11 (tt, J₁=J₂=4.4 Hz, 1H), 1.43 (d, J=6.8 Hz, 3H), 1.36 (t, J=7.72 Hz,3H), 0.84-1.00 (m, 4H) ppm. MS(ESI⁺) 645.3 (MH⁺).

Synthesis of Compound 3.70

Compound 3.70 was synthesized following the synthetic scheme for thegeneric synthesis of 8-azaquinazolinones (FIG. 13) to yield a colorlesssolid. ¹H NMR (d₆-DMSO; T=120° C.) δ8.09 (d, J=7.6 Hz, 1H), 7.87 (dd,J₁=J₂=6.8 Hz, 1H), 7.82 (s, 1H), 7.73 (d, J=8.4 Hz, 1H), 7.56 (dd,J₁=J₂=7.2 Hz, 1H), 7.38 (d, J=6.8 Hz, 2H), 7.30 (dd, J₁=J₂=10.4 Hz, 1H),7.22 (d, J=8.0 Hz, 1H), 7.22 (d, J=8.0 Hz, 1H), 7.08 (d, J=7.6 Hz, 1H),7.03 (d, J=7.6 Hz, 1H), 6.21 (d, J=8.4 Hz, 1H), 5.23 (q, J=6.4 Hz, 1H),4.48 (br s, 2H), 4.09 (q, J=6.8 Hz, 2H), 3.54 (d, J=15.2 Hz, 1H), 3.30(br s, 1H), 2.88 (br s, 4H), 1.91-1.94 (m, 4H), 1.44 (d, J=6.4 Hz, 3H),1.34 (t, J=6.8 Hz, 3H) ppm. MS(ESI⁺) 674.3 (MH⁺).

Synthesis of Compound 3.71

Compound 3.71 was prepared like compound 3.16a, with the pyridyl sidechain prepared from 2,5-dibromopyridine. White solid. ¹H NMR (DMSO, 120°C.) δ8.10 (d, 1H, J=8.0 Hz), 7.86 (m, 2H), 7.73 (d, 1H, J=8.0 Hz), 7.56(dd, 1H, J₁=J₂=8.0 Hz), 7.38 (d, 3H, J=6.8 Hz), 7.31 (d, 1H, J=10.4 Hz),7.25 (d, 1H, J=8.4 Hz), 7.18 (s, 1H), 7.08 (m, 2H), 7.04 (m, 1H), 6.41(d, 1H, J=8.4 Hz), 5.23 (broad s, 1H), 4.49 (s, 2H), 4.12 (q, 1H, J=8.0Hz) 4.09 (q, 2H, J=7.5 Hz), 3.54 (d, 1H, J=13.2 Hz), 2.95 (s, 6H), 1.44(d, 3H, J=6.4 Hz), 1.36 (t, 3H, J=8.0 Hz), 1.28 (s, 1H) ppm. MS (ESI⁺):expected 648.26 (MH⁺), found 648.3.

Synthesis of Compound 3.72

White solid. ¹H NMR (DMSO, 120° C.) δ8.13 (d, 1H, J=8.0 Hz), 7.85 (dd,1H, J₁=J₂=7.6 Hz), 7.67 (d, 1H, J=8.0 Hz), 7.55 (dd, 1H, J₁=J₂=7.6 Hz),7.44 (m, 2H), 7.34 (d, 1H, J=12.4 Hz), 7.30 (dd, 1H, J₁=J₂=9.6 Hz), 7.23(d, 1H, J=8.0 Hz), 7.05 (m, 2H), 5.66 (broad s, 1H), 5.19 (broad s, 1H),4.08 (q, 2H, J=6.6 Hz), 3.77 (hept, 1H, J=6.6 Hz), 3.66 (m, 2H), 3.31(m, 1H), 3.01 (m, 2H), 2.94 (m, 1H), 1.49 (d, 3H, J=6.8 Hz), 1.43 (tq,2H, J=7.2 Hz), 1.35 (t, 3H, J=6.8 Hz), 1.21 (t, 3H, J=7.0 Hz), 1.17 (s,1H), 1.09 (d, 3H, J=6.8 Hz), 0.99 (d, 3H, J=5.2 Hz), 0.83 (t, 3H, J=7.4Hz) ppm. MS (ESI⁺): expected 698.34 (MH⁺), found 698.3.

Synthesis of Compound 3.73

Compound 3.73 was synthesized in the usual fashion, with the pyridylfragment coming from 2,6-dibromopynidine. White solid. ¹H NMR (DMSO,120° C.) δ8.09 (d, 1H, J=8.0 Hz), 7.83 (d, 1H, J₁=J₂=7.8 Hz), 7.68 (d,1H, J=8.0 Hz), 7.53 (dd, 1H, J₁=J₂=7.6 Hz), 7.40 (d, 2H, J=6.4 Hz), 7.27(m, 3H), 7.15 (d, 1H, J₁=₂=7.6 Hz), 7.03 (m, 1H), 6.98 (m, 1H), 6.24 (d,2H, J=7.6 Hz), 5.91 (broad s, 1H), 5.35 (s, 1H), 4.56 (q, 2H, J=16.4Hz), 4.08 (d, 2H, J=6.8 Hz), 3.80 (s, 2H), 2.94 (s, 3H), 2.72 (s, 3H),1.35 (t, 3H, J=6.8 Hz) ppm. MS (ESI⁺): expected 634.25 (MH⁺), found634.2.

Synthesis of Compound 3.74

Compound 3.74 was synthesized as shown in FIG. 13, starting with theL-alanine derivative rather than the D-Ala. White solid. ¹H NMR (DMSO,120° C.) δ9.02 (d, 1H, J=3.6 Hz), 8.46 (dd, 1H, J₁=7.8 Hz, J₂=1.8 Hz),8.35 (s, 2H), 7.57 (dd, 1H, J₁=8.0 Hz, J₂=4.4 Hz), 7.53 (d, 1H, J=6.8Hz), 7.43 (d, 1H, J=6.8 Hz), 7.14 (broad m, 8H), 5.29 (d, 1H, J=6.0 Hz),4.76 (s, 2H), 4.13 (q, 2H, J=6.8 Hz), 3.46 (broad s, 1H), 2.91 (s, 4H),1.42 (d, 3H, J=6.8 Hz), 1.36 (t, 3H, J=7.0 Hz) ppm. MS (ESI⁺): expected604.22 (MH⁺), found 604.3.

Example 4 Synthesis of Compound 4.01

The synthesis of compound 4.01 in four steps from commercially availablestarting materials provides another example of a 3H-quinazolin-4-onesynthesis in racemic form. Scheme 12 provides an overview of thesynthetic route, for which the experimental details follow.

(R)-t-Butyl 2-(N-2-Ethoxyethyl)aminopropionate (XXII). To a solution ofD-alanine t-butyl ester hydrochloride (3.15 g, 17.3 mmol, 1.0 equiv),and 2-bromoethyl ethyl ether (2.79 g, 18.2 mmol, 1.05 equiv) in 14 mL ofDMF, was added KI (1.44 g, 8.7 mmol, 0.50 equiv), followed by K₂CO₃(2.40 g, 17.3 mmol, 1.0 equiv). After stirred at 55° C. for 16 h, thereaction mixture was poured into a mixture of 70 mL of water and 10 mLof 10% Na₂CO₃. The resulting mixture was extracted three times with 50mL of EtOAc. The organic layer was washed with 50 mL of brine, driedover Na₂SO₄ and concentrated in vacuo to give a yellow oil, which waspassed through a short silica gel column, eluted with EtOAc. The eluentwas concentrated in vacuo to give 3.13 g of the crude XXII as a brownoil, which was used in subsequent step without further purification. ¹HNMR (CDCl₃) δ1.20 (t, J=8.0 Hz, 3H), 1.27 (d, J=7.2 Hz, 3H), 1.46 (s,9H), 1.96 (b, 1H), 2.65 (m, 1H), 2.83 (m, 1H), 3.23 (q, J=7.2 Hz, 1H),3.40-3.56 (m, 4H) ppm. MS (ESI⁺) m/z 218.1 [M+H]⁺.

(R)-t-Butyl 2-(N-2-Ethoxyethyl)-(N4-phenylphenylacetyl)aminopropionate(XXIII) To a solution of crude XXII (5.0 g, 23 mmol, 1.0 equiv), and4-phenylphenylacetic acid (4.88 g, 23 mmol, 1.0 equiv) in 40 mL ofdichloromethane, was added EDC (5.51 g, 29 mmol, 1.25 equiv), HOBT (3.89g, 29 mmol, 1.25 equiv), and N-methylmorpholine (2.79 g, 28 mmol, 1.2equiv) at room temperature. The mixture was stirred at room temperaturefor 4 h. The reaction mixture was poured into a 30 mL of 5% aqueousH₃PO₄, and extracted twice with 20 mL of EtOAc. The combined EtOAcextract was washed twice with 20 mL of 10% aqueous NaHCO₃, and once with30 mL of brine. The organic layer was dried over Na₂SO₄ and evaporatedin vacuo to give a brown oil, which was purified by silica gelchromatography to give 5.05 g of compound XXIII as a light yellow oil.¹H NMR (CDCl₃) δ1.20 (t, J=8.0 Hz, 3H), 1.27 (d, J=7.2 Hz, 3H), 1.46 (s,9H), 1.95 (br, 1H), 2.65 (m, 1H), 2.83 (m, 1H), 3.23 (q, J=7.2 Hz, 3H),3.56 (m, 4H) ppm. MS (ESI⁺) m/z 218.1 [M+H]⁺.

(R)-2-(N-2-Ethoxyethyl)-(N-4-phenylphenylacetyl)aminopropionic acid(XXIV) To a solution of compound XXIII (5.05 g, 12.3 mmol, 1.0 equiv) in25 mL of dichloromethane, was added triethylsilane (3.57 g, 30.7 mmol,2.5 equiv), and trifluoroacetic acid (18 g, 160 mmol, 13 equiv) at roomtemperature. The mixture was stirred at room temperature for 8 h. Thereaction mixture was evaporated in vacuo to give a brown residue, whichwas dissolved in 60 mL of EtOAc and washed once with 50 mL of 0.5 Maqueous KH₂PO₄, followed by 40 mL of brine. The organic layer was driedover Na₂SO₄ and evaporated in vacuo to give a brown oil, which waspurified by silica gel chromatography to give 3.69 g of compound XXIV asa colorless oil, which solidified into a cream colored solid uponstanding at room temperature. At room temperature the product exists asmixture of cis/trans amide rotamers, ca. 4.4:1 molar ratio in DMSO. Forthe major rotamer, ¹H NMR (DMSO-d₆) δ1.12 (t, J=7.0 Hz, 3H), 1.34 (d,J=6.8 Hz, 3H3.40-3.60 (m, 6H), 3.78 (s, 2H), 4.16 (q, J=6.8 Hz, 1H),7.29 (d, J=8.0 Hz, 2H), 7.35 (t, J=7.2 Hz, 1H), 7.46 (t, J=7.4 Hz, 2H),7.59 (d, J=8.0 Hz, 2H), 7.65 (d, J=7.8 Hz, 2H) ppm. For the minorrotamer ¹H NMR (DMSO-d₆) δ4.77 (q, J=6.8 Hz, 1H) ppm. MS (ESI⁻) m/z354.2 [M−H]⁻.

2-((N-2-Ethoxyethyl)-N-(4-phenylphenylacetyl)-1-aminoethyl)-3-(6-benzothiazolyl)-3H-quinazoline-4-one(4.01). To a solution of anthranilic acid (69 mg, 0.50 mmol, 1.0 equiv)and compound XXIV (178 mg, 0.50 mmol, 1.0 equiv) in 1.0 mL of anhydrouspyridine was added 127 μL of triphenylphosphite (155 mg, 0.50 mmol, 1.0equiv) at room temperature. The resulting yellow solution was stirred atreflux for 2 h. 6-Aminobenzothiazole (75 mg, 0.50 mmol, 1.0 equiv) wasadded via syringe. The reaction mixture was stirred for another 3 h at100° C., cooled to room temperature, and evaporated in vacuo to give abrown residue. This residue was dissolved in 20 mL of ether. The mixturewas washed successively twice with 5 mL of 5% aqueous phosphoric acid,twice with 5 mL of 1 M NaOH, once with 5 mL of pH 7 phosphate buffer(0.5 M KH₂PO₄ and 0.5 M K₂HPO₄), and once with 10 mL of brine. Theorganic layer was dried over Na₂SO₄ and evaporated in vacuo to give abrown residue, which was purified by preparative TLC to give 19 mg ofcompound 4.01 as a light yellow solid. At room temperature, thiscompound exists as a mixture of cis/trans amide rotamers, anddiastereomers, ca. 0.33:0.30:1 molar ratio in DMSO. ¹H NMR (DMSO-d₆,T=25° C.) δ4.92 (q, J=6.8 Hz, 1H), 5.05 (q, J=6.8 Hz, 1H), & 5.27 (q,J=6.8 Hz, 1H) ppm. MS (ESI⁺) m/z 589.3 [M+H]⁺.

Synthesis of Compound 4.03

Compound 4.03 was prepared following the synthesis of compound 4.01.Yellow solid, mixture of cis/trans amide rotamers (1.5/1), determined by¹H NMR (CDCl₃) 1.20 (t, 3H, J=7.0 Hz), 1.26 (t, 3H, J=7.0 Hz). MS(ESI⁺)577.3 (MH⁺).

Example 5 Synthesis of Compound 5.01

Synthesis of the biphenyl compound 5.01 was achieved via a four-stepreaction sequence, commencing with a Suzuki coupling of1-ethyl-2-iodo-benzene and 4-ethoxyphenylboronic acid to form thebiphenyl unit. The remaining transformations install the amino alkyl andacetyl groups.

4′-Ethoxy-2-ethyl-biphenyl. A degassed (3×freeze-thaw cycles) mixture of1.00 mL 1-ethyl-2-iodo-benzene (6.97 mmol, 1.00 equiv), 3.47 g4-ethoxyphenylboronic acid (20.9 mmol, 3.00 equiv), and 402 mgtetrakis(triphenylphosphine)palladium(0) (0.349 mmol, 0.0501 equiv) wasdissolved in 8.0 mL toluene and 8.0 mL aqueous 2M sodium carbonatesolution and the biphasic mixture heated to 100° C. (externaltemperature, oil bath). After 16 h the reaction was cooled to roomtemperature and the organic phase separated. The aqueous layer wasextracted with 50% ethyl acetate in hexane (2×25 mL) and the combinedorganic separations dried over magnesium sulfate, filtered, andconcentrated in vacuo to yield a yellow oil. The crude material waspurified by column chromatography on silica gel (3.5 cm o.d.×20 cm h)eluting with 5% ethyl acetate in hexane. Fractions containing product atR_(f)=0.68, 10% ethyl acetate in hexane, were combined and concentratedin vacuo to afford 1.54 g product, including impurity, as a colorlessoil; ca. 1.23 g pure product. An impurity of ca. 20%, identified as thehomocoupling product 4,4′-diethoxybiphenyl and quantified by relativeratio of integrated ¹H NMR resonance signals, was carried forward withthe product to the next step. ¹H NMR (CDCl₃) δ1.15 (t, 3H, J=7.6 Hz),1.49 (t, 3H, J=7.2 Hz), 2.65 (q, 2H, J=7.6 Hz), 4.12 (q, 2H, J=7.2 Hz),6.98 (d, 2H, J=8.4 Hz), 7.22-7.28 (m, 2H), 7.27 (d, 2H, J=8.4 Hz),7.31-7.34 (m, 2H) ppm.

2-(1-Bromo-ethyl)-4′-ethoxy-biphenyl from 4′-ethoxy-2-ethyl-biphenyl. Amixture of 673 mg 4′-ethoxy-2-ethyl-biphenyl (2.98 mmol, 1.00 equiv),556 mg N-bromosuccinimide (3.13 mmol, 1.05 equiv), and 49 mg2,2′-azobisisobutyronitrile (0.30 mmol, 0.10 equiv) dissolved in 15 mLcarbon tetrachloride was heated to reflux in the presence of a highintensity incandescent light for 1.5 h. The reaction was cooled to 0° C.and the resulting precipitate removed by filtration. The concentratedfiltrate was subjected to iterative triturations with cold hexane (3×50mL) to afford 890 mg product as a colorless oil. The4,4′-diethoxybiphenyl impurity, ca. 20% quantitated by relative ratio ofintegrated ¹H NMR resonance signals, was carried forward to the nextstep with the product. ¹H NMR (CDCl₃) δ1.48 (t, 3H, J=6.8 Hz), 1.99 (d,3H, J=7.2 Hz), 4.12 (q, 2H, J=6.8 Hz), 5.33 (q, 1H, J=6.8 Hz), 7.00 (d,2H, J=8.8 Hz), 7.21 (d, 1H, J=7.6 Hz), 7.30-7.34 (m, 3H), 7.42 (dd, 1H,J₁=J₂=7.6 Hz), 7.77 (d, 1H, J=8.0 Hz)

[1-(4′-Ethoxy-biphenyl-2-yl)-ethyl]-(2-ethoxy-ethyl)-amine from2-(1-bromo-ethyl-4′-ethoxy-biphenyl. A mixture of 135 mg2-(1-bromo-ethyl)-4′-ethoxy-biphenyl (0.442 mmol, 1.00 equiv) and 115 μL2-ethoxy-1-aminoethane (1.10 mmol, 2.50 equiv) dissolved in 3.0 mLethanol was heated to reflux for 20 h and then concentrated in vacuo toremove the solvent. The concentrated reaction product was adsorbeddirectly onto a column of silica gel (3.5 cm o.d.×12 cm h) and elutedwith 3% methanol in chloroform. Fractions containing product atR_(f)=0.30, 10% methanol in chloroform, were combined and concentratedin vacuo to afford 13.5 mg purified product as a yellow oil. ¹H NMR(CDCl₃) δ1.17 (t, 3H, J=7.2 Hz), 1.34 (d, 3H, J=6.8 Hz), 1.47 (t, 3H,J=7.72 Hz,), 2.55 (t, 2H, J=5.6 Hz), 3.37-3.50 (m, 2H), 4.01 (q, 1H,J=6.4 Hz), 4.10 (q, 2H, J=6.8 Hz), 6.94 (d, 2H, J=8.8 Hz), 7.15-7.22 (m,3H), 7.27 (dd, 1H, J₁=J₂=7.6 Hz), 7.39 (dd, 1H, J₁=J₂=7.6 Hz), 7.63 (d,1H, J=7.6 Hz) ppm. MS (ESI, positive mode) 314.1 [MH]⁺.

A mixture of 13.5 mg[1-(4′-ethoxy-biphenyl-2-yl)-ethyl]-(2-ethoxy-ethyl)-amine (43.1 μmol,1.00 equiv), 10.5 mg 4′-(trifluoromethyl)phenylacetic acid (51.7 μmol,1.20 equiv), 9.9 mg EDC (51.7 μmol, 1.20 equiv), and 1.0 mg HOBT (7.4μmol, 0.18 equiv) dissolved in 2.0 mL dichloromethane was stirred atroom temperature for 2 h. To the reaction solution was added 5 mLaqueous saturated sodium bicarbonate solution. The aqueous layer wasdiluted with water to 15 mL and extracted with dichloromethane (2×20mL). The combined organic separations were dried over magnesium sulfate,filtered, and concentrated in vacuo to yield a yellow oil. The crudeproduct was adsorbed onto a column of silica gel (3.5 cm o.d.×10 cm h)and eluted with 17% to 25% ethyl acetate gradient in hexane. Fractionscontaining product were combined and concentrated in vacuo to afford12.8 mg purified product as a colorless oil. ¹H NMR (d₆-DMSO; T=140° C.)δ1.04 (t, 3H, J=6.8 Hz), 1.34 (t, 3H, J=6.8 Hz), 1.44 (d, 3H, J=7.2 Hz),3.00-3.06 (m, 1H), 3.17-3.36 (m, 6H), 3.49 (d, 1H, J=16.0 Hz), 4.07 (q,2H, J=6.8 Hz), 5.43 (q, 1H, J=7.2 Hz), 6.92-6.96 (m, 2H), 7.13-7.18 (m,3H), 7.22-7.26 (m, 2H), 7.34 (ddd, 1H, J₁=1.2 Hz, J₂=7.6 Hz, J₃=8.4 Hz),7.39 (ddd, 1H, J₁=2.0 Hz, J₂=7.6 Hz, J₃=9.2 Hz), 7.53-7.59 (m, 3H) ppm.At room temperature, compound exists as a mixture of cis/trans amiderotamers in ca. 2:1 ratio as determined by integration of characteristic¹H NMR signals (CDCl₃; T=25° C.) δ_(major) 5.19 (q, 2.1H, J=7.2 Hz) andδ_(minor) 5.89 (q, 1.0H, J=7.6 Hz) ppm. MS (ESI, positive mode) 500.1[MH]⁺

Example 6 Synthesis of Compound 6.01

A mixture of compound 3.22 (13 mg) and ammonium acetate (500 mg) inacetic acid (2 mL) was stirred at 80° C. for 14 h and at 100° C. for 10h. The acetic aid was evaporated, and the residue was taken by EtOAc. Itwas washed with sodium bicarbonate and brine, dried, and concentrated.The residue was purified by column (70% EtOAc in Hexane) to give 10 mgof compound 6.01. ¹H NMR (CDCl₃) δ8.26 (d, J=8.0 Hz, 1H), 7.84 (m, 2H),7.56 (m, 1H), 7.46 (m, 2H), 7.32 (m, 2H), 7.18 (m, 1H), 7.05 (m, 2H),6.60 (s, 1H), 6.53 (m, 1H), 4.90 (q, 1H), 3.76 (d, 1H), 2.62(d, 1H),2.16 (s, 3H), 1.27 (d, 3H). MS (ESI⁺) 507.2 [MH]⁺.

Synthesis of Compound 6.02

Into a mixture of bromide (0.557 mmol, 0.20 g) and K₂CO₃ (0.89 mmol,0.123 g) in 3 mL of DMF was added 2-undecyl-1-H-imidazole (0.557 mmol,0.124 g). The reaction mixture was heated to 90° C. for 10 h. Afterevaporating the solvent, the residue was dissolved in CH₂Cl₂, theorganic layer was washed by water, brine, dried over NaSO₄ and removedin vacuo to give a sticky oil which was purified by chromatography toafford a yellow solid (0.16 g). ¹H NMR (CDCl₃) 0.89 (t, 3H, J=7.0 Hz),1.25 (m, 16H), 1.60 (br m, 3H), 1.73 (d, 3H, J=6.7 Hz), 1.83 (m, 1H),3.84 (s, 3H), 5.09 (q, 1H, J=6.7 Hz), 6.35 (m, 1H), 6.84-6.90 (m, 3H),7.05 (m, 1H), 7.20 (m, 1H), 7.55 (m, 1H), 7.82 (m, 2H), 8.28 (dd, 1H,J₁=1.1 Hz, J₂=7.9 Hz). MS(ESI⁺) 501.2 (MH⁺). Anal. (C₃₁H₄₀N₄O₂) cal. C,74.22; H, 8.05; N, 11.19. Found C, 74.22; H, 8.14; N, 11.03.

Preparation of Compound 6.03

The synthesis of compound 6.03 is shown in FIG. 16. ¹H NMR (CDCl₃) δ8.27(d, J=8.0 Hz, 1H), 7.80 (m, 2H), 7.74 (s, 1H), 7.57 (m, 1H), 7.48 (m,2H), 7.21 (m, 2H), 7.02 (m, 3H), 6.64 (m, 1H), 5.10 (q, J=6.8 Hz, 1H),4.12 (q, 2H), 3.83 (d, J=16.4 Hz, 1H), 2.95 (d, J=16.2 Hz, 1H), 1.49 (t,3H), 1.32 (d, J=6.7 Hz, 3H). MS (ESI⁺) 544.2 [MH]⁺.

Preparation of Compound 6.04

The synthesis of compound 6.04 is shown in FIG. 16. ¹H NMR (CDCl₃) δ8.26(d, J=8.0 Hz, 1H), 7.80 (m, 2H), 7.54 (m, 1H), 7.47 (m, 2H), 7.18 (m,2H), 7.03 (m, 2H), 6.96 (m, 1H), 6.87 (s, 1H), 6.56 (m, 1H), 5.01 (q,J=6.8 Hz, 1H), 4.11 (q, 2H), 3.71 (d, J=16.4 Hz, 1H), 2.83 (m, 2H), 2.76(d, J=16.2 Hz, 1H), 2.69 (m, 2H), 1.47 (t, 3H), 1.27 (d, J=6.7 Hz, 3H).MS (ESI⁺) 572.3 [MH]⁺.

Preparation of Compound 6.05

The synthesis of compound 6.05 is shown in FIG. 16. ¹H NMR (CDCl₃) δ8.26(d, J=8.0 Hz, 1H), 7.80 (m, 2H), 7.54 (m, 1H), 7.47 (m, 2H), 7.18 (m,2H), 7.06 (m, 2H), 6.96 (m, 1H), 6.78 (s, 1H), 6.52 (m, 1H), 5.01 (q,J=6.8 Hz, 1H), 4.11 (m, 3H), 3.64 (m, 2H), 3.34 (s, 3H), 2.81 (m, 2H),2.68 (d, J=16.2 Hz, 1H), 1.47 (t, 3H) 1.24 (d, J=6.7 Hz, 3H). MS (ESI⁺)577.1 [MH]⁺.

Preparation of Compound 6.06

The synthesis of compound 6.06 is shown in FIG. 16. ¹H NMR (CDCl₃) δ8.36(m, 1H), 8.27 (d, J=8.0 Hz, 1H), 7.80 (m, 2H), 7.56 (m, 1H), 7.47 (m,3H), 7.16 (m, 2H), 7.04 (m, 3H), 6.92 (m, 2H), 6.50 (s, 1H), 6.29 (m,1H), 4.92 (q, J=6.8 Hz, 1H), 4.11 (q, 2H), 3.74 (d, J=16.4 Hz, 1H), 3.08(m, 2H), 2.98 (m, 2H), 2.59 (d, J=16.2 Hz, 1H), 1.47 (t, 3H), 1.17 (d,J=6.7 Hz, 3H). MS (ESI⁺) 624.2 [MH]⁺.

Preparation of Compound 6.07

The synthesis of compound 6.07 is shown in FIG. 16. ¹H NMR (CDCl₃) δ8.26(d, J=8.0 Hz, 1H), 7.80 (m, 2H), 7.54 (m, 1H), 7.47 (m, 2H), 7.18 (m,2H), 7.05 (m, 2H), 6.96 (m, 1H), 6.72 (s, 1H), 6.48 (m, 1H), 5.00 (q,J=6.8 Hz, 1H), 4.11 (q, 2H), 4.04 (q, 2H), 3.80 (d, J=16.4 Hz, 1H), 2.85(m, 2H), 2.65 (m, 3H), 1.47 (t, 3H), 1.23 (d, J=6.7 Hz, 3H), 1.14 (t,3H). MS (ESI⁺) 619.1 [MH]⁺.

Preparation of Compound 6.08

The synthesis of 6.08 is shown in FIG. 16. ¹H NMR (CDCl₃) δ8.53 (m, 1H),8.26 (d, J=8.0 Hz, 1H), 7.80 (m, 2H), 7.50 (m, 5H), 7.26 (m, 5H), 7.07(m, 3H), 7.00 (m, 1H), 6.61 (m, 1H), 5.03 (q, J=6.8 Hz, 1H), 4.11 (q,2H), 3.89 (d, J=16.14 Hz, 1H), 2.81 (d, J=16.2 Hz, 1H), 1.47 (t, 3H),1.27 (d, J=6.7 Hz, 3H). MS (ESI⁺[MH]⁺.

Preparation of Compound 6.09

The synthesis of compound 6.09 is shown in FIG. 16. ¹H NMR (CDCl₃) δ8.27(d, J=8.0 Hz, 1H), 7.80 (m, 2H), 7.53 (m, 1H), 7.48 (m, 3H), 7.25 (s,1H), 7.20 (m, 2H), 7.05 (m, 2H), 7.00 (m, 1H), 6.58 (m, 1H), 6.50 (d,J=15.8 Hz, 1H), 5.04 (q, J=6.8 Hz, 1H), 4.21 (q, 2H), 4.12 (q, 2H), 3.83(d, J=16.4 Hz, 1H), 2.80 (d, J=16.2 Hz, 1H), 1.48 (t, 3H), 1.28 (d,J=6.7 Hz, 3H). MS (ESI⁺) 617.2 [MH]⁺.

Preparation of Compound 6.10

The synthesis of compound 6.10 is shown in FIG. 16. ¹H NMR (CDCl₃) δ8.26(d, J=8.0 Hz, 1H), 7.80 (m, 2H), 7.54 (m, 1H), 7.47 (m, 2H), 7.19 (m,2H), 7.04 (m, 3H), 6.96 (m, 1H), 6.59 (m, 1H), 5.04 (q, J=6.8 Hz, 1H),4.44 (s, 2H), 4.11 (m, 3H), 3.60 (m, 2H), 2.70 (d, J=16.2 Hz, 1H), 1.47(t, 3H), 1.24 (d, J=6.7 Hz, 3H). MS (ESI⁺) 577.5 [MH]⁺.

Preparation of Compound 6.11

The synthesis of compound 6.11 is shown in FIG. 16. ¹H NMR (CDCl₃) δ8.27(d, J=8.0 Hz, 1H), 7.80 (m, 2H), 7.54 (m, 1H), 7.46 (m, 2H), 7.19 (m,2H), 7.04 (m, 2H), 6.97 (m, 2H), 6.54 (m, 1H), 5.03 (q, J=6.8 Hz, 1H),4.53 (s, 2H), 4.11 (q, 2H), 3.75 (d, J=16.4 Hz, 1H), 2.73 (d, J=16.2 Hz,1H), 1.47 (t, 3H), 1.27 (d, J=6.7 Hz, 3H). MS (ESI⁺) 549.5 [MH]⁺.

Example 7

Compound 7.01. To a solution of the amine (1 mmol, 0.37 g) anddiisopropylethylamine (1.2 mmol, 0.16 g) in acetonitrile (3 mL) andmethylene chloride (3 mL) was added octanesulfonyl chloride (1.2 mmol,0.26 g). The reaction mixture was stirred at room temperature overnight.Sodium carbonate (15%) was added and the aqueous layer was extractedwith methylene chloride. The organic layer was washed with water, brine,dried over NaSO₄ and concentrated in vacuo to give a yellow oil, whichwas purified by chromatography on silica gel (eluent: CHCl₃/MeOH=10/1.5)to afford a light yellow glassy oil (0.22 g). ¹H NMR (CDCl₃) 0.86 (t,3H, J=7.2 Hz), 1.17-1.25 (m, 10H), 1.46 (d, 3H, J=6.9 Hz), 1.70 (m, 2H),2.19 (s, 6H), 2.45 (m, 2H), 2.73-2.90 (m, 2H), 3.62-3.78 (m, 2H), 3.87(s, 3H), 4.88(q, 1H, J=6.9 Hz), 7.04-7.14 (m, 3H), 7.31-7.34 (m, 1H),7.49 (dt, 1H, J₁=1.3 Hz, J₂=8 Hz), 7.68 (d, 1H, J=7.3 Hz), 7.77 (dd, 1H,J₁=2.1 Hz, J₂=8 Hz), 8.27 (dd, 1H, J₁=1.2 Hz, J₂=8 Hz). MS(ESI⁺) 544.2(MH⁺). Anal. (C₂₉H₂N₄O₄S) cal. C, 64.18; H, 7.80; N, 10.32; S, 5.91.Found C, 64.36; H, 7.81; N, 10.08; S, 5.78

Example 8

Compound 8.01. Compound 8.01 was prepared using the similar conditionfor synthesis of compound 4.01. Oil. Mixture of cis/trans amide rotamers(1/6), determined by ¹H NMR (CDCl₃) 4.18 (q, 1H, J=7.0 Hz), 4.69 (q, 1H,J=7.0 Hz). MS(ESI⁺) 533.2 (MH⁺).

Example 9

2-Amino-N-(4-ethoxy-phenyl)benzamide (XX). A mixture of isotoicanhydride (16.3 g, 100 mmol) and p-phenetidine (13.7 g, 100 mmol) washeated at 120° C. for 4 h. The reaction mixture after cooling wastriturated with ether. The resulting solid was collected by suction togive compound XX, ¹H NMR (CD₃OD) 1.37 (t, 3H, J=7.0 Hz), 4.01 (q, 2H,J=7.0 Hz), 6.67 (t, 1H, J=7.0 Hz), 6.78 (dd, 1H, J₁=1.2 Hz, J₂=8.2 Hz),6.89 (m, 2H), 7.20 (dt, 1H, J₁=1.4 Hz, J₂=8.2 Hz), 7.47 (m, 2H), 7.56(dd, 1H, J₁=1.4 Hz, J₂=9.3 Hz).MS(ESI⁺) 257.3 (MH⁺).

Synthesis of o-diamide XXI. To a mixture of compound XX (7.68 g, 30mmol) and N-(9-fluorenylmethyloxycarbonyl)-D-alanine (10.26 g, 33 mmol)in CH₂Cl₂ (150 mL), was added EDAC (8.63 g, 45 mmol) and HOBt (1.38 g, 9mmol). After stirring at room temperature overnight, the resulting solidwas filtered and washed with ethyl ether to yield compound XXI (14.50g). ¹H NMR (CDCl₃) 1.37 (t, 3H, J=7.0 Hz), 1.48 (d, 3H, J=7.2 Hz), 3.89(m, 2H), 4.26 (m, 2H), 4.45 (m, 2H), 5.50 (m, 1H), 6.76 (m, 2H), 7.17(t, 1H, J=7.3 Hz), 7.25-7.76 (m, 12H), 8.62 (d, 1H, J=8.8 Hz), 11.43 (s,1H).MS(ESI⁺) 550.3 (MH⁺).

4-Oxoquinazoline XXII. To a solution of diamide XXI (7.27 g, 13.27 mmol)in CH₂Cl₂, was added PPh₃ (17.40 g, 66.39 mmol), I₂ (16.52 g, 65.02mmol) and N,N-diisopropylethylamine(17.12 g, 132.7 mmol). The reactionmixture was stirred at room temperature overnight. The resulting solidwas filtered and washed with ethyl ether to yield compound XXII (4.83g). ¹H NMR (CDCl₃) 1.43 (t, 3H, J=7.0 Hz), 1.52 (d, 3H, J=7.2 Hz), 4.03(m, 2H), 4.23 (m, 1H), 4.43 (m, 2H), 4.66 (m, 1H), 5.58 (m, 2H), 6.88(m, 2H), 7.23-7.78 (m, 13H), 8.46 (d, 1H, J=8.8 Hz).MS(ESI⁺) 532.3(MH⁺).

Compound XXIII. Piperidine (15 ml) was added to a solution of compoundXXII (2.68 g, 5.05 mmol) in DMF (100 ml). After stirring at roomtemperature for 1 h, the mixture was poured into 150 ml of water, theaqueous layer was extracted with CH₂Cl₂, the combined organic extractswas dried over Na₂SO₄, filtered and concentrated. The residue waspurified by chromatography to give a white solid (0.80 g). ¹H NMR(CDCl₃) 1.30 (d, 3H, J=6.6 Hz), 1.46 (t, 3H, J=6.3 Hz), 3.82 (m, 1H),4.10 (q, 2H, J=6.6 Hz), 7.03 (dd, 2H, J₁=1.9 Hz, J₂=7.0 Hz), 7.18 (m,2H), 7.47 (m, 1H), 7.75 (m, 2H), 8.26 (d, 1H, J=8 Hz). MS(ESI⁺) 310.1(MH⁺).

Compound XXIV. To a mixture of compound XXIII (0.06 g, 0.19 mmol) andbromoacetamide (0.032 g, 0.23 mmol) in DMF (3 mL), was added K₂CO₃(0.079 g, 0.57 mmol) and NaI (0.086 g, 0.57 mmol). After stirring atroom temperature overnight, evaporated the solvent, the residue wasdissolved in CH₂Cl₂, the organic layer was washed by water, brine, driedover NaSO₄ and removed in vacuo to give a yellow solid which waspurified by chromatography to afford a white solid. ¹H NMR (CDCl₃) 1.26(t, 3H, J=7.2 Hz), 1.38(d, 3H, J=6.6 Hz), 3.46 9br, 1H), 3.58 (br, 1H),3.82 (m, 1H), 4.11 (m, 2H), 5.68 (br, 1H), 7.04 (m, 2H), 7.14 (m, 2H),7.50 (m, 1H), 7.75 (m, 2H), 8.30 (d, 1H, J=8 Hz). MS(ESI⁺) 367.3 (MH⁺).

Synthesis of Compound 9.01

Compound 9.01 was prepared using the similar condition for synthesis ofcompound 3.02, white solid, mixture of cis/trans amide rotamers (1/5),determined by ¹H NMR (CDCl₃) 4.85 (q, 1H, J=7.3 Hz), 5.35 (q, 1H, J=7.3Hz). MS(ESI⁺) 561.2

Synthesis of Compound 9.02

Compound 9.02 was prepared following the synthesis of compound 9.01,oil, mixture of cis/trans amide rotamers (1/1), determined by ¹H NMR(CDCl₃) 4.95 (m, 1H), 5.35 (m, 1H). MS(ESI⁺) 630.2 (MH⁺).

Synthesis of Compound 9.03

Compound 9.03 was prepared following the synthesis of compound 9.01.Yellow solid, m.p. 167.9° C., mixture of cis/trans amide rotamers (1/2),determined by ¹H NMR (CDCl₃) 4.85 (q, 1H, J=7.0 Hz), 5.26 (q, 1H, J=7.0Hz). MS(ESI⁺) 604.2 (MH⁺). Anal. (C₃₄H₂₉F₄N₃O₃) cal. C, 67.65; H, 4.84;N, 6.96. Found C, 67.80; H, 4.98; N, 6.97.

Synthesis of Compound 9.04

Compound 9.04 was prepared following the synthesis of compound 9.01.white solid, m.p. 156.2° C. ¹H NMR (DMSO, T=140° C.) 1.45(d, 3H, J=6.8Hz), 3.59-3.73 (m, 6H), 3.92 (m, 2H), 5.14(q, 1H, J=6.8 Hz), 7.33 (m,5H), 7.69 (m, 4H), 7.72 (d, 1H, J=8 Hz), 7.86 (m 1H), 8.14 (dd, 1H,J=1.2 Hz, J=8.4 Hz), At room temperature, mixture of cis/trans amiderotamers (1/2), determined by ¹H NMR (CDCl₃) 4.88 (q, 1H, J=6.8 Hz),5.27 (q, 1H, J=6.8 Hz). MS(ESI⁺) 604.2 (MH⁺). Anal. (C₂₉H₂₄F₇N₃O₃) cal.C, 58.49; H, 4.06; N, 7.06. Found C, 58.53; H, 4.18; N, 7.05.

Synthesis of Compound 9.05

Compound 9.05 was prepared following the synthesis of compound 9.01.Yellow solid, mixture of cis/trans amide rotamers (1/1), determined by¹H NMR (CDCl₃) 4.88 (q, 1H, J=7.0 Hz), 5.35 (q, 1H, J=7.0 Hz). MS(ESI⁺)518.3 (MH⁺). Anal. (C₃₀H₃₂FN₃O₄) cal. C, 69.62; H, 6.23; N, 8.12. FoundC, 69.40; H, 6.26; N, 7.98.

Synthesis of Compound 9.06

Compound 9.06 was prepared following the synthesis of compound 9.01.Yellow solid, mixture of cis/trans amide rotamers (1/1), determined by¹H NMR (CDCl₃) 4.90 (q, 1H, J=7.0 Hz), 5.35 (q, 1H, J=7.0 Hz). MS(ESI⁺)518.3 (MH⁺). Anal. (C₃₀H₃₂ FN₃O₄) cal. C, 69.62; H, 6.23; N, 8.12. FoundC, 69.33; H, 6.20; N, 8.06.

Synthesis of Compound 9.07

Compound 9.07 was prepared following the synthesis of compound 9.01.Yellow solid, mixture of cis/trans amide rotamers (1/1), determined by¹H NMR (CDCl₃) 4.88 (q, 1H, J=7.0 Hz), 5.37 (q, 1H, J=7.0 Hz). MS(ESI⁺)536.3 (MH⁺). Anal. (C₃₀H₃₁F₂N₃O₄) cal. C, 67.28; H, 5.83; N, 7.85. FoundC, 67.28; H, 5.80; N, 7.78.

Synthesis of Compound 9.08

Compound 9.08 was prepared following the synthesis of compound 9.01.Yellow solid, m.p. 157.9° C. ¹H NMR (DMSO, T=140° C.) 0.95 (t, 3H, J=6.4Hz), 1.34 (t, 3H, J=6.8 Hz), 1.44 (d, 3H, J=6.8 Hz), 3.31-3.59 (m, 8H),4.08 (q, 2H, J=6.8 Hz), 5.17 (q, 1H, J=6.8 Hz), 7.02 (m, 2H), 7.24-7.56(m, 7H), 7.70 (d, 1H, J=8 Hz), 7.84 (dt, 1H, J₁=1.6 Hz, J₂=7.2 Hz), 8.13(d, 1H, J=8 Hz). At room temperature, mixture of cis/trans amiderotamers (1/1), determined by ¹H NMR (CDCl₃) 4.92 (q, 1H, J=7.0 Hz),5.38 (q, 1H, J=7.0 Hz). MS(ESI⁺) 568.3 (MH⁺). Anal. (C₃₁H₃₂ F₃N₃O₄) cal.C, 65.60; H, 5.68; N, 7.40. Found C, 65.38; H, 5.61; N, 7.34.

Synthesis of Compound 9.09

Compound 9.09 was prepared following the synthesis of compound 9.01.Colorless oil, ¹H NMR (CDCl₃) 1.04 (t, 3H, J=6.9 Hz), 1.46 (m, 6H), 3.30(m, 2H), 3.42 (m, 2H), 3.62 (m, 2H), 4.08 (q, 2H, J=7.0 Hz), 5.15 (q,1H, J=7.0 Hz), 7.02 (m, 2H), 7.18 (m, 1H), 7.42-7.54 (m, 8H), 7.75 (m,1H), 8.28 (d, 1H, J=7.8 Hz) MS(ESI⁺) 536.3 (MH⁺).

Example 10 Synthesis of Compound 10.01

A mixture of compound XV (160 mg, 0.5 mmol) and 2-imidazolcarboxaldehyde(58 mg, 0.6 mmol) in methanol (10 mL) was stirred at room temperaturefor 20 minutes. Then sodium cyanoborohydride (38 mg, 0.6 mmol) wasadded. The mixture was stirred at room temperature for 6 h. The reactionmixture was treated with EtOAc, and it was washed with sodiumbicarbonate and brine, dried, and concentrated. The residue was purifiedby column (5% methanol and 1% conc. NH₄OH in 3:7 EtOAc/DCM) to give 120mg of compound XXV. ¹H NMR (CDCl₃) δ8.25 (d, J=8.0 Hz, 1H), 7.78 (t,J=8.0 Hz, 1H), 7.70 (d, J=7.7 Hz, 1H), 7.49 (t, J=8.0 Hz, 1H), 7.20-7.05(m, 4H), 6.93 (s, 2H), 3.94 (d, J=14.8 Hz, 1H), 3.77 (d, J=14.8 Hz, 1H),3.38 (q, J=6.6 Hz, 1H), 1.25 (d, J=6.6 Hz, 3H).

EDC (123 mg, 0.64 mmol) was added to a mixture of compound XXV (115 mg,0.32 mmol), 4-trifluoromethylphenyl acetic acid (65 mg, 0.32 mmol), HOBt(43 mg, 0.32 mmol), and NMM (0.07 mL, 0.64 mmol) in DMF (3 mL). Themixture was stirred at room temperature for 14 h. The reaction mixturewas treated with EtOAc, and it was washed with sodium bicarbonate andbrine, dried, and concentrated. The residue was purified by column (5%methanol and 1% conc. NH₄OH in 3:7 EtOAc/DCM) to give 100 mg of compound10.01. MS (ESI⁺) 550.2 [MH]⁺.

Synthesis of Compound 10.02

Potassium carbonate (97 mg, 0.7 mmol) was added to a mixture of compound10.01 (38 mg, 0.07 mmol) and iodomethane (0.044 mL, 0.7 mmol) in DMF (2mL). The mixture was stirred at room temperature for two days. DMF wasevaporated under high vacuum, and the residue was taken by EtOAc. It waswashed with brine, dried, and concentrated. The residue was purified bycolumn (2% methanol and 0.5% conc. NH₄OH in 3:7 EtOAc/DCM) to give 15 mgof compound 10.02. MS (ESI⁺) 564.2 [MH]⁺.

Synthesis of 10.03

Compound 10.03 was prepared following the synthetic procedure forcompound 10.01, described above. MS (ESI⁺) 550.2 [MH]⁺.

Synthesis of Compound 10.04

Compound 10.04 was prepared following the synthetic procedure forcompound 10.02, described above. MS (ESI⁺) 564.2 [MH]⁺.

Synthesis of Compound 10.05

Compound 10.05 was prepared following the synthetic procedure ofcompound 10.01, described above. ¹H NMR (d₆-DMSO, T=140° C.) δ8.11 (d,J=8.0 Hz, 1H), 7.82 (t, J=8.1 Hz, 1H), 7.68 (d, J=8.1 Hz, 1H), 7.60-7.40(m, 6H), 7.38-7.15 (m, 5H), 5.33 (bs, 1H), 5.02 (dd, J=11.4 Hz, 2H),3.60 (bm, 2H), 1.45 (d, J=7.0 Hz, 3H). m.p. 173-174° C. MS (ESI⁺) 567.2[MH]⁺. Anal. Calcd. for C₂₉H₂₂F₄N₄O₂S; C, 61.48; H, 3.91; N, 9.89.Found: C, 61.36; H, 4.08; N, 9.75.

Example 11 Preparation of Compound 11.01

The synthesis of compound 11.01 is shown in FIG. 14. ¹H NMR (d₆-DMSO,T=120° C.) δ8.8-7.0 (m, 17H), 6.35 (s, 1H), 5.00 (m, 1H), 4.35 (m, 1H),4.25-4.00 (m, 4H), 3.65 (m, 1H), 1.40 (t, 3H). MS (ESI⁺) 619.1 [MH]⁺.

Preparation of Compound 11.02

The synthesis of compound 11.02 is shown in FIG. 17. ¹H NMR (d₆-DMSO,T=150° C.) δ8.38 (m, 2H), 8.09 (m, 1H), 7.84 (m, 2H), 7.68 (m, 1H), 7.54(m, 2H), 7.40-7.00 (m, 6H), 5.25 (m, 1H), 4.74 (m, 2H), 4.25 (m, 1H),4.14 (m, 3H), 3.62 (m, 1H), 3.31 (m, 1H), 2.78 (m, 2H), 1.37 (m, 6H). MS(ESI⁺) 605.3 [MH]⁺.

Preparation of Compound 11.03

The synthesis of compound 11.03 is shown in FIG. 18. ¹H NMR (d₆-DMSO,T=150° C.) δ9.02 (s, 1H), 8.85 (s, 1H), 8.40 (s, 1H), 8.38 (m, 1H), 7.57(m, 1H), 7.41 (m, 3H), 7.33-7.15 (m, 5H), 5.30 (q, 1H), 4.80 (dd, 2H),4.13 (q, 2H), 3.64 (d, 1H), 3.21 (bs, 1H), 1.46 (d, 3H), 1.37 (t, 3H).MS (ESI⁺) 607.2 [MH]⁺.

Preparation of Compound 11.04

The synthesis of compound 11.04 is shown in FIG. 18. ¹H NMR (d₆-DMSO,T=150° C.) δ8.81 (d, 1H), 8.38 (s, 1H), 8.35 (m, 1H), 8.03 (m, 1H), 7.81(m, 1H), 7.54 (m, 1H), 7.40 (m, 3H), 7.33-7.05 (m, 5H), 5.28 (q, 1H),4.75 (dd, 2H), 4.13 (q, 2H), 3.60 (d, 1H), 3.19 (bs, 1H), 1.44 (d, 3H),1.37 (t, 3H). MS (ESI⁺) 606.2 [MH]⁺.

Preparation of Compound 11.05

The synthesis of compound 11.05 is shown in FIG. 11. ¹H NMR (d₆-DMSO,T=150° C.) δ8.47 (s, 1H), 8.37 (d, 1H), 8.13 (d, 1H), 7.66-7.25 (m, 7H),7.25-7.10 (m, 6H), 6.59 (d, 1H), 5.25 (q, 1H), 4.86 (dd, 2H), 4.18 (q,2H), 3.70 (d, 1H), 3.31 (bd, 1H), 1.41 (m, 6H). MS (ESI⁺) 605.2 [MH]⁺.

Preparation of Compound 11.06

The synthesis of compound 11.06 is shown in FIG. 3. ¹H NMR (d₆-DMSO,T=150° C.) δ8.30 (m, 1H), 8.22 (s, 1H), 7.95 (m, 2H), 7.83 (m, 1H), 7.57(m, 1H), 7.48 (m, 1H), 7.36 (m, 2H), 7.24 (m, 2H), 7.15-6.95 (m, 6H),5.45 (q, 1H), 4.50 (dd, 2H), 4.14 (q, 2H), 3.57 (d, 1H), 3.05 (bd, 1H),1.54 (d, 3H), 1.38 (t, 3H). MS (ESI⁺) 587.3 [MH]⁺.

Preparation of Compound 11.07

The synthesis of compound 11.07 is shown in FIG. 1. ¹H NMR (CDCl₃) δ8.30(m, 3H), 7.85 (m, 2H), 7.52 (m, 3H), 7.23 (m, 1H), 7.11 (m, 1H), 7.03(m, 2H), 6.82 (d, 1H), 6.75 (d, 1H), 6.54 (m, 1H), 5.07 (q, 1H), 4.60(dd, 2H), 4.05 (m, 3H), 3.82 (m, 2H), 1.85 (d, 1H), 1.45 (t, 3H), 1.17(s, 9H). MS (ESI⁺) 677.3 [MH]⁺.

Example 12

This example illustrates a CXCR3 binding assay that can be used forevaluating the compounds of the present invention.

Unless otherwise noted, all reagents used are available from commercialsources (e.g., Sigma). Test compounds are diluted in DMSO to aconcentration that is 40 times the intended final assay concentration; 5μL are transferred to each well of a 96-well flat-bottomed polypropyleneplate (e.g., from Greiner, Inc.). CXCR3-expressing cells obtained fromChemoCentryx were used in the assays to generate the data set forth inthe Table provided in FIG. 12. The cells were resuspended in assaybuffer (25 mM Hepes, 80 mM NaCl, 1 mM CaCl₂, 5 mM MgCl₂, 0.2% bovineserum albumin, pH 7.1, stored at 4° C.) at 5 million cells per mL; 100μL of this cell suspension is then transferred to each well of a 96-wellplate containing the diluted test compounds. ¹²⁵I-labelled chemokine(purchased from commercial sources, e.g., Amersham, PE Life Sciences) isdiluted in assay buffer to a concentration of approximately 60 pM; 100μL of this chemokine solution is transferred to each well of a 96-wellplate containing compounds and cell suspension. The plates are sealedwith commercially available foil plate seals (e.g., from E&KScientific), and stored at 4° C. for 2 to 4 h, shaking gently. At theend of this incubation period, the contents of the assay plates aretransferred to GF/B filter plates (Packard) that have been pre-coated bydipping into a solution containing 0.3% polyethyleneimine (Sigma), usinga cell harvester (Packard), and washing twice with wash buffer (25 mMHepes, 500 mM NaCl, 1 mM CaCl₂, 5 mM MgCl₂, pH 7.1, stored at roomtemperature). The filter plates are sealed on the bottom with plateseals (Packard), 50 μL of Microscint-20 scintillation fluid (Packard) isadded to each well, and the top of the plates are sealed with clearplastic (TopSeal A, Packard). The plates are counted on a scintillationcounter, such as a Packard TopCount. To measure non-specific binding, 4wells containing unlabelled “cold” chemokine were included on each96-well plate. To measure maximum binding, 4 wells containing 5 μL ofDMSO, 100 μL of cell suspension and 100 μL of ¹²⁵I-labelled chemokinesolution were included on each 96-well plate. Data were analyzed usingcommercially available software (e.g., Excel from Microsoft, Prism fromGraphPad Software Inc.).

Other assays may be used to identify compounds that modulate CXCR3chemokine receptor activity, for example, binding assays (see, e.g.,Weng et al. (1998) J. Biol. Chem. 273:18288-18291, Campbell et al.(1998) J. Cell Biol. 141:1053-1059, Endres et al. (1999) J. Exp. Med.189:1993-1998 and Ng et al. (1999) J. Med. Chem. 42:4680-4694), calciumflux assays (see, e.g., Wang et al. (2000) Mol. Pharm. 57:1190-1198 andRabin et al. (1999) J. Immunol. 162:3840-3850) and chemotaxis assays(see, e.g., Albanesi et al. (2000) J. Immunol. 165:1395-1402 andLoetscher et al. (1998) Eur. J. Immunol. 28:3696-3705).

All publications and patent applications cited in this specification areherein incorporated by reference as if each individual publication orpatent application were specifically and individually indicated to beincorporated by reference. Although the foregoing invention has beendescribed in some detail by way of illustration and example for purposesof clarity of understanding, it will be readily apparent to those ofordinary skill in the art in light of the teachings of this inventionthat certain changes and modifications may be made thereto withoutdeparting from the spirit or scope of the appended claims.

What is claimed is:
 1. A compound having the formula:

or a pharmaceutically acceptable salt, isomer or prodrug thereof.
 2. Thecompound of claim 1 or a pharmaceutically acceptable salt thereof. 3.The compound of claim
 1. 4. A pharmaceutical composition comprising thecompound of claim 1 and a pharmaceutically acceptable carrier ordiluent.
 5. A method of treating an inflammatory or immune condition ordisease in a subject, said method comprising administering to a subjectin need of such treatment a therapeutically effective amount of thecompound of claim
 1. 6. The method of claim 5, wherein said compound isadministered topically.
 7. The method of claim 5, wherein said compoundmodulates CXCR3.
 8. The method of claim 5, wherein said inflammatory orimmune condition or disease is selected from the group consisting ofneurodegenerative diseases, multiple sclerosis, systemic lupuserythematosus, rheumatoid arthritis, atherosclerosis, encephalitis,meningitis, hepatitis, nephritis, sepsis, sarcoidosis, psoriasis,eczema, uticaria, type I diabetes, asthma, conjunctivitis, otitis,allergic rhinitis, chronic obstructive pulmonary disease, sinusitis,dermatitis, inflammatory bowel disease, Behcet's syndrome, gout, viralinfections, bacterial infections, organ transplant conditions and skintransplant conditions.
 9. The method of claim 8 wherein saidinflammatory bowel disease is ulcerative colitis or Crohn's disease. 10.The method of claim 5, wherein said compound is administered incombination with a second therapeutic agent, wherein said secondtherapeutic agent is useful for treating neurodegenerative diseases,multiple sclerosis, systemic lupus erythematosus, rheumatoid arthritis,atherosclerosis, encephalitis, meningitis, hepatitis, nephritis, sepsis,sarcoidosis, psoriasis, eczema, uticaria, type I diabetes, asthma,conjunctivitis, otitis, allergic rhinitis, chronic obstructive pulmonarydisease, sinusitis, dermatitis, inflammatory bowel disease, Behcet'ssyndrome, gout, viral infections, bacterial infections, organ transplantconditions or skin transplant conditions.
 11. The method of claim 10wherein said inflammatory bowel disease is ulcerative colitis or Crohn'sdisease.
 12. A method of treating a CXCR3-mediated condition or diseasein a subject, said method comprising administering to a subject in needof such treatment a therapeutically effective amount of the compound ofclaim
 1. 13. A method in accordance with claim 12, wherein saidCXCR3-mediated condition is selected from the group consisting ofneurodegenerative diseases, multiple sclerosis, systemic lupuserythematosus, rheumatoid arthritis, atherosclerosis, encephalitis,meningitis, hepatitis, nephritis, sepsis, sarcoidosis, psoriasis,eczema, uticaria, type I diabetes, asthma, conjunctivitis, otitis,allergic rhinitis, chronic obstructive pulmonary disease, sinusitis,dermatitis, inflammatory bowel disease, Behcet's syndrome, gout, viralinfections, bacterial infections, organ transplant conditions and skintransplant conditions.
 14. The method of claim 13 wherein saidinflammatory bowel disease is ulcerative colitis or Crohn's disease. 15.The method of claim 12, wherein said compound modulates CXCR3.
 16. Themethod of claim 12, wherein said compound is administered in combinationwith a second therapeutic agent, wherein said second therapeutic agentis useful for treating neurodegenerative diseases, multiple sclerosis,systemic lupus erythematosus, rheumatoid arthritis, atherosclerosis,encephalitis, meningitis, hepatitis, nephritis, sepsis, sarcoidosis,psoriasis, eczema, uticaria, type I diabetes, asthma, conjunctivitis,otitis, allergic rhinitis, chronic obstructive pulmonary disease,sinusitis, dermatitis, inflammatory bowel disease, Behcet's syndrome,gout, viral infections, bacterial infections, organ transplantconditions or skin transplant conditions.
 17. The method of claim 16wherein said inflammatory bowel disease is ulcerative colitis or Crohn'sdisease.
 18. The method of claim 16, wherein said organ transplantcondition is a bone marrow transplant condition or a solid organtransplant condition.
 19. The method of claim 18, wherein said solidorgan transplant condition is a kidney transplant condition, a livertransplant condition, a lung transplant condition, a heart transplantcondition or a pancreas transplant condition.
 20. A method in accordancewith claim 12, wherein said CXCR3-mediated condition is psoriasis.
 21. Amethod in accordance with claim 12, wherein said CXCR3-mediatedcondition is inflammatory bowel disease.
 22. A method in accordance withclaim 12, wherein said CXCR3-mediated condition is selected from thegroup consisting of multiple sclerosis, rheumatoid arthritis and organtransplant conditions.
 23. A method in accordance with claim 12, whereinsaid compound is used in conjunction with another therapeutic agentselected from the group consisting of Remicade®, Enbrel®, a COX-2inhibitor, a glucocoiticoid, an immunosuppressant, methotrexate,prednisolone, azathioprine, cyclophosphamide, tacrolimus, mycophenolate,hydroxychloroquine, sulfasalazine, cyclosponne A, D-penicillaniine, agold compound, an antilymphocyte or antithymocyte globulin, betaseron,avonex and copaxone. 24.A method in accordance with claim 12, whereinsaid CXCR3-mediated condition is an organ transplant condition and saidcompound is used alone or in combination with a second therapeutic agentselected from the group consisting of cyclosporine A, FK-506, rapamycin,mycophenolate, prednisolone, azathioprine, cyclophosphamide and anantilymphocyte globulin.
 25. A method in accordance with claim 12,wherein said CXCR3-mediated condition is rheumatoid arthritis and saidcompound is used alone or in combination with a second therapeutic agentselected from the group consisting of methotrexate, sulfasalazine,hydroxychloroquine, cyclosporine A, D-penicillamine, Remicade®, Enbrel®,auranofin and aurothioglucose.
 26. A method in accordance with claim 12,wherein said CXCR3-mcdiated condition is multiple sclerosis and saidcompound is used alone or in combination with a second therapeutic agentselected from the group consisting of betaseron, avonex, azathioprine,copaxone, prednisolone and cyclophosphamide.
 27. A method in accordancewith claim 11, wherein said subject is a human.
 28. A method for themodulation of CXCR3 function in a cell, comprising contacting said cellwith the compound of claim
 1. 29. A method for the modulation of CXCR3function, comprising contacting a CXCR3 protein with the compound ofclaim
 1. 30. A method of treating asthma in a subject, said methodcomprising administering to a subject in need of such treatment atherapeutically effective amount of the compound of claim 1 or a saltthereof.
 31. A method of treating rheumatoid arthritis in a subject,said method comprising administering to a subject in need of suchtreatment a therapeutically effective amount of the compound of claim 1or a salt thereof.
 32. A method of treating psoriasis in a subject, saidmethod comprising administering to a subject in need of such treatment atherapeutically effective amount of the compound of claim 1 or a saltthereof.
 33. A method of treating inflammatory bowel disease in asubject, said method comprising administering to a subject in need ofsuch treatment a therapeutically effective amount of the compound ofclaim 1 or a salt thereof.
 34. A method of treating multiple sclerosisin a subject, said method comprising administering to a subject in needof such treatment a therapeutically effective amount of the compound ofclaim 1 or a salt thereof.
 35. A method of treating organ transplantconditions in a subject, said method comprising administering to asubject in need of such treatment a therapeutically effective amount ofthe compound of claim 1 or a salt thereof.
 36. The pharmaceuticalcomposition of claim 4 in a form suitable for topical use.
 37. Thepharmaceutical composition of claim 4 in a form of a sterile aqueous oroleagenous solution or suspension suitable for parenteral injection. 38.The pharmaceutical composition of claim 4 in a form suitable for oraluse.
 39. The method of claim 5, wherein said compound is administeredparenterally.
 40. The method of claim 5, wherein said compound isadministered orally.